In vitro bactericidal assay Fig. S8 Gentamicin protection assay Phagocytosis assay

Size: px
Start display at page:

Download "In vitro bactericidal assay Fig. S8 Gentamicin protection assay Phagocytosis assay"

Transcription

1 In vitro bactericidal assay Mouse bone marrow was isolated from the femur and the tibia. Cells were suspended in phosphate buffered saline containing.5% BSA and 2 mm EDTA and filtered through a cell strainer. Red blood cells were removed and neutrophils were enriched by negative selection using an EasySep system (StemCell Technologies, Vancouver, BC, Canada). We routinely obtain 4-5 million cells per mouse, and >9% of them are morphologically mature neutrophils (Fig. S8). Purified neutrophils were treated with different concentrations of SF167 or DMSO (vehicle control) for 3 min at 37 C then washed twice with HBSS. Live E. coli particles (strain 19138; American Type Culture Collection, cfu) were opsonized with mouse serum (final concentration 12.5%) at 37 C for 3 min and then incubated with pretreated neutrophils ( cfu) at 37 C for 1 hour. Reactions were stopped and neutrophils were lysed with sterile water (1:1). Samples were then serially diluted and spread on Luria broth (LB) agar plates. Colony numbers were determined after overnight incubation at 37 C. Gentamicin protection assay Neutrophil intracellular bactericidal activity was measured using a gentamicin protection assay. Briefly, mouse bone marrow derived neutrophils were treated with SF167 or DMSO (vehicle control) for 3 min at 37 C then washed twice with HBSS. Cells were incubated with serum-opsonized live E. coli particles (as described above) at a ratio of 1:1 at 37 C for 1 hour. Gentamicin was then added at the final concentration of 1 g/ml to kill extracellular bacteria. After additional 1 hour incubation, cells were washed twice with HBSS and lysed with 1 ml sterile water at room temperature. Viable intracellular bacteria were quantified by subsequent plating the lysed samples on LB agar plates. The results were normalized by phagocytosis indexes. Phagocytosis assay Fluorescein-conjugated Zymosan bioparticles (Molecular Probes) were reconstituted in HBSS and opsonized with 12.5% mouse serum at 37 C for 3 min. Mouse bone marrow derived neutrophils were incubated with serum-opsonized bioparticles at a ratio of 1:1 (neutrophils : bioparticles) at 37 C for 1 hour. Negative controls were incubated on ice instead. The fluorescence of extracellular particles was 1

2 quenched by addition of 1 μl of.25 mg/ml trypan blue. The number of internalized particles was counted under a fluorescence microscope (Olympus lx17, 1X objective). Phagocytosis efficiency was expressed as the number of the internalized particles per 1 neutrophils (Phagocytosis index). Binding efficiency was expressed as the number of the binding particles per 1 neutrophils (Binding index). More than 1 cells were counted from random fields per coverslip for each group. Bacterial burden and BALF total protein levels Bronco alveolar lavage (BAL) samples were collected from mice at 24 hr after bacteria challenge. For bacterial burden assay, 1 ml of cold PBS/15 mm EDTA was injected to the lung and withdrew while gently massaging lungs. This was repeated with 1 ml PBS/15 mm EDTA buffer. BALF were then pooled and serially diluted with ice-cold sterile PBS. Aliquots of 1 l were spread on LB agar plates (E.coli), chocolate II agar (S. pneumonia), or Columbia agar with 5% sheep blood (S. aureus). Colonies were counted after overnight incubation. For BALF total protein assay, BAL was done with 1 ml of ice cold PBS/15 mm EDTA. Protein concentration was measured using the Bio-Rad protein assay reagent. The standard curve was constructed using BSA. Histopathology Lungs were collected 24 hr after infection. Whole lungs were inflated and fixed with Bouin s solution. The specimens were embedded in paraffin and 6 m thick sections were stained with hematoxylin and eosin and examined by light microscopy. Nonquantitative histological analysis was performed by a pathologist blinded for groups. IPLab software (BD Biosciences) was used to manually trace edema containing regions in the tissue section. The pixel area of each edema containing region was calculated using the same IPLab software ( area measurement modular). The edema formation was calculated as the percentage of pixel area of all the edema containing regions relative over the pixel area of the whole image. 2

3 3 min 16 hr DMSO DMSO SF: 125 nm 7AAD SF: 25 nm 7AAD SF: 25 nm SF: 5 nm Annexin V Annexin V Figure S1. Treatment with SF167 does not significantly affect neutrophil spontaneous death. Human neutrophils (1 x 1 6 ) were treated with indicated concentrations of SF167 at 37 C for 3 min and washed twice with PBS. After being cultured in RPMI164 plus 1% FBS for indicated time, the amounts of healthy and apoptotic cells were determined by AnnexinV-APC and 7-Amino-actinomycin D (7-AAD) staining as previously described (Zhu et al., 26).

4 A fmlp 1µM fmlp 1µM ionomycin B [Ca2 2+] ( F34/38) ) [Ca2 2+] ( F34/38) ) F34/38 Figure S2. Treatment with SF167 does not alter fmlp-induced calcium signaling in neutrophils. Mouse bone marrow-derived neutrophils were pretreated with or without SF167 at 37 C for 3 min, and then loaded with 1 µm of Fluo-4-AM and 2 µm of Fura-Red AM in 1 ml PBS at RT for 45 min. Cells were washed hdtwice with ihpbs and resuspended ddi in (A) HBSS (no calcium, no magnesium) + 1% FBS + 2 nm HEPES + 1 mm EDTA or (B) HBSS (with calcium and magnesium) + 1% FBS + 2 nm HEPES. Cells were allowed to settle at least 15 min at RT before being stimulated with 1 M of fmlp. The changes in intracellular calcium ([Ca 2 ]i) were monitored for 5 min after stimulation. Black line, DMSO vehicle control; blue line, 5 nm of SF167; brown line, 25 nm of SF167; green line,125 nm of SF167. Arrows, addition of fmlp or ionomycin. The signal detected in (A) in the absence of extracellular calcium represents calcium release from intracellular calcium stores.

5 DMSO control DMSO control DMSO 3nM FITC-fMLP 6nM FITC-fMLP 3nM FITC-fMLP + 1 M fmlp 6nM FITC-fMLP + 1M fmlp control 12nM FITC-fMLP 12nM FITC-fMLP + 1M fmlp Events SF:5nM control 3nM FITC-fMLP 3nM FITC-fMLP + 1 M fmlp SF:5nM control 6nM FITC-fMLP 6nM FITC-fMLP + 1 M fmlp SF:5nM control 12nM FITC-fMLP 12nM FITC-fMLP + 1 M fmlp FITC Figure S3. Treatment with SF167 does not change surface expression of fmlp receptors in neutrophils. Human neutrophils (.5 x 1 6 ) treated with or without 5 nm of SF167 (37 C for 3 min) were pre-blocked on ice for 45 min in HBSS supplemented with 1% BSA/.2% sodium azide. Cells were then incubated with 3 nm, 6 nm, or 12 nm of FITC-fMLP (Invitrogen, Carlsbad, CA) in the same medium (for 1 hour on ice), in the presence or absence of 1 M of unlabeled fmlp, washed once with HBSS, and immediately analyzed by flow cytometry. Neutrophils were gated by forward/side scatter. Specific binding of FITC-fMLP was determined by subtracting geometric mean fluorescence intensity in the presence of 1 M unlabeled fmlp from the geometric mean fluorescence intensity in the absence of 1 M unlabeled fmlp.

6 DMSO SF:1nM SF:1nM SF:1nM Figure S4. PTEN inhibitor SF167 does not have bactericidal activity. Live E. coli particles (strain 19138; American Type Culture Collection, cfu) were incubated with indicated concentrations of SF167 in HBSS at 37 C for 1 hour (total volume 1 µl). Samples were then diluted and spread on LB agar plates. Colonies in each sample were counted after overnight incubation at 37 C.

7 A Neutropenic mice Recipient i mice Induction of neutropenia with Cy Intraperitoneal injection of ftg(3%) (3%), 2.5 hours Mixed (1:1) population was iv injected into the inflamed mice Donor mice CMTMR labeling -1.5 hours after neutrophil transfusion, collect cells from peritoneal cavity. CMTM MR CMTMR+ Input control Bone marrow derived d neutrophils 1.5h after iv injection CMFDA labeling B C D CMTMR+ Relativ ve amount of neutr rophils (CMDMR/CMFDA A) Input control Peritoneal lavage - Determine the ratio of (CMTMR + cells)/(cmfda )( + cells) by FACS analysis. red blood cells Neutrophils lymphocytes CMFDA Figure S5. Recruitment of transfused neutrophils to the inflamed peritoneal cavity. (A) Schematic representation of the experimental system for neutrophil transfusion. Neutrophils isolated from wild-type mice were treated with or without SF167 for 3 min at 37 C, washed twice with PBS, and then labeled with CMTMR or CMFDA. Labeled cells were mixed (1:1) as indicated and then transfused intravenously (via tail vein) to neutropenic mice that had been challenged with 3% thioglycollate (TG) for 2.5 hr. Peritoneal lavage was harvested 1.5 hr after the neutrophil transfusion. (B) Labeling with fluorescent dye only does not affect neutrophil recruitment. Bone marrow derived neutrophils were labeled with CMFDA or CMTMR. The amount of adoptively transferred neutrophils recruited to the peritoneal cavity were analyzed using a FACSCanto II flow cytometer and FACSDiva software. (C) Relative recruitment of neutrophils was calculated as the ratio of indicated populations in the peritoneal lavage. Data shown are mean ± SD of three mice. (D) Forward and side scatter shows that most of the non-neutrophil populations in the inflamed peritoneal cavity are red blood cells with small amount of lymphocytes.

8 Cy: 15 mg/kg ip 1mg/kg ip Bacteria challenge & Neutrophil transfusion Days Peripheral blood neutrophils (x1million n/ml) d1 d2 d3 d4 d5 d6 d7 WT Figure S6. Schematic of mouse neutropenia model. Mouse neutropenia was induced by two intraperitoneal injections of cyclophosphamide (Cy) at a total dose of 25 mg/kg (15 mg/kg on day 1 and 1 mg/kg on day 4). On day 5, Cy-treated mice contained approximately 9% fewer circulating neutrophils than untreated or saline-treated group. The profound neutropenia persisted through days 6 and 7.

9 DMSO control SF167 Arbitrary Lig ght Units (-) fmlp (+) fmlp t Units Arbirary Ligh (-) fmlp (+) fmlp Time (sec) Time (sec) TNFa priming TNFa priming + SF167 trary Light Units Arbit 5 (-) fmlp 4 (+) fmlp Time (sec) Arbitra ary Light Units 5 (-) fmlp 4 (+) fmlp Time (sec) Figure S7. PTEN inhibitor SF167 can further enhance ROS production in mouse neutrophils primed with TNFa. Bone marrow derived mouse neutrophils were treated with TNFa (5ng/mL), SF167 (5nM), TNFa+SF167, or DMSO alone at 37ºC for 3 min in HBSS (.2% BSA) buffer, washed twice with HBSS, and then stimulated with HBSS or 2 µm of fmlp. ROS production was monitored in the presence of 5 M of isoluminol,.8u of HRP in a luminometer at 37 C. Chemiluminescence (arbitrary light units) was recorded at indicated time points.

10 Figure S8. The purity of isolated mouse neutrophils. Mouse bone marrow was isolated from the femur and the tibia. Cells were suspended in phosphate buffered saline containing.5% BSA and 2 mm EDTA and filtered through a cell strainer. Red blood cells were removed and neutrophils were enriched by negative selection using an EasySep system (StemCell Technologies, Vancouver, BC, Canada). We routinely obtain 4-5 million cells per mouse, and >9% of them are mature neutrophils.

Supplementary Figures

Supplementary Figures Inhibition of Pulmonary Anti Bacterial Defense by IFN γ During Recovery from Influenza Infection By Keer Sun and Dennis W. Metzger Supplementary Figures d a Ly6G Percentage survival f 1 75 5 1 25 1 5 1

More information

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk

Procaspase-3. Cleaved caspase-3. actin. Cytochrome C (10 M) Z-VAD-fmk. Procaspase-3. Cleaved caspase-3. actin. Z-VAD-fmk A HeLa actin - + + - - + Cytochrome C (1 M) Z-VAD-fmk PMN - + + - - + actin Cytochrome C (1 M) Z-VAD-fmk Figure S1. (A) Pan-caspase inhibitor z-vad-fmk inhibits cytochrome c- mediated procaspase-3 cleavage.

More information

Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham

Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham Primary Adult Naïve CD4+ CD45RA+ Cells Prepared by: David Randolph (drdrdr@uab.edu) at University of Alabama, Birmingham Goal: To obtain large numbers of highly pure primary CD4+ CD45RO- CD25- cells from

More information

Gladstone Institutes, University of California (UCSF), San Francisco, USA

Gladstone Institutes, University of California (UCSF), San Francisco, USA Fluorescence-linked Antigen Quantification (FLAQ) Assay for Fast Quantification of HIV-1 p24 Gag Marianne Gesner, Mekhala Maiti, Robert Grant and Marielle Cavrois * Gladstone Institutes, University of

More information

The Annexin V Apoptosis Assay

The Annexin V Apoptosis Assay The Annexin V Apoptosis Assay Development of the Annexin V Apoptosis Assay: 1990 Andree at al. found that a protein, Vascular Anticoagulant α, bound to phospholipid bilayers in a calcium dependent manner.

More information

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze. This document is available at www.stemcell.com/pis EasySep Mouse Monocyte Isolation Kit Catalog #19861 For processing 1 x 10^9 cells Description Isolate untouched and highly purified monocytes from mouse

More information

10.00 PBS OVA OVA+isotype antibody 8.00 OVA+anti-HMGB1. PBS Methatroline (mg/ml)

10.00 PBS OVA OVA+isotype antibody 8.00 OVA+anti-HMGB1. PBS Methatroline (mg/ml) RESEARCH ARTICLE Penh (100% of PBS) 1 PBS 8.00 +anti-hmgb1 6.00 4.00 p=0.054 Cellular & Molecular Immunology advance online publication, PBS 3.12 6.25 Methatroline (mg/ml) Neutrophil isolation and culture

More information

Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2*

Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2* Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2* 1 Department of Laboratory Medicine - Laboratory of Hematology, Radboud University

More information

Instructions for Use. APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests

Instructions for Use. APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests 3URGXFW,QIRUPDWLRQ Sigma TACS Annexin V Apoptosis Detection Kits Instructions for Use APO-AB Annexin V-Biotin Apoptosis Detection Kit 100 tests For Research Use Only. Not for use in diagnostic procedures.

More information

Figure S1. PMVs from THP-1 cells expose phosphatidylserine and carry actin. A) Flow

Figure S1. PMVs from THP-1 cells expose phosphatidylserine and carry actin. A) Flow SUPPLEMENTARY DATA Supplementary Figure Legends Figure S1. PMVs from THP-1 cells expose phosphatidylserine and carry actin. A) Flow cytometry analysis of PMVs labelled with annexin-v-pe (Guava technologies)

More information

VEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization

VEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization Cell Reports, Volume 9 Supplemental Information VEGFR2-Mediated Vascular Dilation as a Mechanism of VEGF-Induced Anemia and Bone Marrow Cell Mobilization Sharon Lim, Yin Zhang, Danfang Zhang, Fang Chen,

More information

Annexin V-APC/7-AAD Apoptosis Kit

Annexin V-APC/7-AAD Apoptosis Kit Annexin V-APC/7-AAD Apoptosis Kit Catalog Number KA3808 100 assays Version: 04 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4

More information

HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation

HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation SUPPLEMENTARY INFORMATION Materials and Methods Human cell lines and culture conditions HCC1937 is the HCC1937-pcDNA3 cell line, which was derived from a breast cancer with a mutation in exon 20 of BRCA1

More information

CHAPTER 4 IMMUNOLOGICAL TECHNIQUES

CHAPTER 4 IMMUNOLOGICAL TECHNIQUES CHAPTER 4 IMMUNOLOGICAL TECHNIQUES Nitroblue Tetrazolium Chloride (NBT) Reduction test NBT reduction test was evaluated by employing the method described by Hudson and Hay,1989 based upon principle that

More information

Interferon γ regulates idiopathic pneumonia syndrome, a. Th17 + CD4 + T-cell-mediated GvH disease

Interferon γ regulates idiopathic pneumonia syndrome, a. Th17 + CD4 + T-cell-mediated GvH disease Interferon γ regulates idiopathic pneumonia syndrome, a Th17 + CD4 + T-cell-mediated GvH disease Nora Mauermann, Julia Burian, Christophe von Garnier, Stefan Dirnhofer, Davide Germano, Christine Schuett,

More information

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze. This document is available at www.stemcell.com/pis Catalog #18765 EasySep Mouse CD4+CD62L+ T Cell Isolation Kit For processing 1x 10^9 cells Description Isolate highly purified naïve CD4+ T cells (CD4+CD62L+)

More information

T H 1, T H 2 and T H 17 polarization of naïve CD4 + mouse T cells

T H 1, T H 2 and T H 17 polarization of naïve CD4 + mouse T cells A complete workflow for cell preparation, isolation, polarization and analysis T H 1, T H 2 and T H 17 polarization of naïve CD4 + mouse T cells Introduction Workflow CD4 + T helper (T H) cells play a

More information

Protocol for Gene Transfection & Western Blotting

Protocol for Gene Transfection & Western Blotting The schedule and the manual of basic techniques for cell culture Advanced Protocol for Gene Transfection & Western Blotting Schedule Day 1 26/07/2008 Transfection Day 3 28/07/2008 Cell lysis Immunoprecipitation

More information

Islet viability assay and Glucose Stimulated Insulin Secretion assay RT-PCR and Western Blot

Islet viability assay and Glucose Stimulated Insulin Secretion assay RT-PCR and Western Blot Islet viability assay and Glucose Stimulated Insulin Secretion assay Islet cell viability was determined by colorimetric (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay using CellTiter

More information

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze.

COMPONENT NAME COMPONENT # QUANTITY STORAGE SHELF LIFE FORMAT. Store at 2-8 C. Do not freeze. Store at 2-8 C. Do not freeze. This document is available at www.stemcell.com/pis Catalog #18765 EasySep Mouse CD4+CD62L+ T Cell Isolation Kit For processing 1x 10^9 cells Description Isolate highly purified naïve CD4+ T cells (CD4+CD62L+)

More information

Evaluation of Antibacterial Effect of Odor Eliminating Compounds

Evaluation of Antibacterial Effect of Odor Eliminating Compounds Evaluation of Antibacterial Effect of Odor Eliminating Compounds Yuan Zeng, Bingyu Li, Anwar Kalalah, Sang-Jin Suh, and S.S. Ditchkoff Summary Antibiotic activity of ten commercially available odor eliminating

More information

ROS Activity Assay Kit

ROS Activity Assay Kit ROS Activity Assay Kit Catalog Number KA3841 200 assays Version: 03 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information... 4 Materials

More information

Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained

Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained 1 2 3 4 5 6 7 8 9 10 11 Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained jejunum sections ( 200 magnification;

More information

In vitro human regulatory T cell expansion

In vitro human regulatory T cell expansion - 1 - Human CD4 + CD25 + regulatory T cell isolation, Workflow in vitro expansion and analysis In vitro human regulatory T cell expansion Introduction Regulatory T (Treg) cells are a subpopulation of T

More information

In vitro human regulatory T cell expansion

In vitro human regulatory T cell expansion - 1 - Human CD4 + CD25 + CD127 dim/- regulatory T cell Workflow isolation, in vitro expansion and analysis In vitro human regulatory T cell expansion Introduction Regulatory T (Treg) cells are a subpopulation

More information

Supplementary Figure 1. Tamoxifen does not promote chemotaxis or chemokinesis in the absence of additional stimulation. Transwell chemotaxis assays

Supplementary Figure 1. Tamoxifen does not promote chemotaxis or chemokinesis in the absence of additional stimulation. Transwell chemotaxis assays Supplementary Figure 1. Tamoxifen does not promote chemotaxis or chemokinesis in the absence of additional stimulation. Transwell chemotaxis assays revealed that tamoxifen treatment alone did not stimulate

More information

Calibration Protocols

Calibration Protocols Calibration Protocols (1)OD 600 Reference point LUDOX Protocol Materials: 1ml LUDOX CL-X (provided in kit) ddh20 (provided by team) 96 well plate, black with clear flat bottom preferred (provided by team)

More information

Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and

Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and Supplementary Figure S1. Flow cytometric analysis of the expression of Thy1 in NH cells. Flow cytometric analysis of the expression of T1/ST2 and Thy1 in NH cells derived from the lungs of naïve mice.

More information

Comparison of primary tumor sections from MMTV-PyMT or MTLn3-ErbB3-

Comparison of primary tumor sections from MMTV-PyMT or MTLn3-ErbB3- Supplemental Data Comparison of primary tumor sections from MMTV-PyMT or MTLn3-ErbB3- GFP tumors in mice either injected with control or clodronate-containing liposomes and stained for macrophages using

More information

Annexin V-PE Apoptosis Detection Kit

Annexin V-PE Apoptosis Detection Kit Annexin V-PE Apoptosis Detection Kit Catalog Number KA0716 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General Information...

More information

The Schedule and the Manual of Basic Techniques for Cell Culture

The Schedule and the Manual of Basic Techniques for Cell Culture The Schedule and the Manual of Basic Techniques for Cell Culture 1 Materials Calcium Phosphate Transfection Kit: Invitrogen Cat.No.K2780-01 Falcon tube (Cat No.35-2054:12 x 75 mm, 5 ml tube) Cell: 293

More information

Supplementary Materials for

Supplementary Materials for immunology.sciencemag.org/cgi/content/full/2/16/eaan6049/dc1 Supplementary Materials for Enzymatic synthesis of core 2 O-glycans governs the tissue-trafficking potential of memory CD8 + T cells Jossef

More information

Modulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis

Modulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis icell Skeletal Myoblasts Application Protocol Modulating Glucose Uptake in Skeletal Myotubes: Insulin Induction with Bioluminescent Glucose Uptake Analysis Introduction The skeletal muscle is one of the

More information

LDL Uptake Flow Cytometry Assay Kit

LDL Uptake Flow Cytometry Assay Kit LDL Uptake Flow Cytometry Assay Kit Item No. 601470 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION

More information

SUPPLEMENTARY INFORMATION. CXCR4 inhibitors could benefit to HER2 but not to Triple-Negative. breast cancer patients

SUPPLEMENTARY INFORMATION. CXCR4 inhibitors could benefit to HER2 but not to Triple-Negative. breast cancer patients SUPPLEMENTARY INFORMATION CXCR4 inhibitors could benefit to HER2 but not to Triple-Negative breast cancer patients Lefort S. 1,2, Thuleau A. 3, Kieffer Y. 1,2, Sirven P. 1,2, Bieche I. 4, Marangoni E.

More information

Supplementary Figure 1 Lymphocytes can be tracked for at least 4 weeks after

Supplementary Figure 1 Lymphocytes can be tracked for at least 4 weeks after Supplementary Figure 1 Lymphocytes can be tracked for at least 4 weeks after photoconversion by using H2B-Dendra2. 4-5 PPs of H2B-Dendra2 BM chimeras were photoconverted and analyzed 7 days (upper panel)

More information

Effective Targeting of Quiescent Chronic Myelogenous

Effective Targeting of Quiescent Chronic Myelogenous Cancer Cell, Volume 7 Supplemental Information Effective Targeting of Quiescent Chronic Myelogenous Leukemia Stem Cells by Histone Deacetylase Inhibitors in Combination with Imatinib Mesylate Bin Zhang,

More information

TFEB-mediated increase in peripheral lysosomes regulates. Store Operated Calcium Entry

TFEB-mediated increase in peripheral lysosomes regulates. Store Operated Calcium Entry TFEB-mediated increase in peripheral lysosomes regulates Store Operated Calcium Entry Luigi Sbano, Massimo Bonora, Saverio Marchi, Federica Baldassari, Diego L. Medina, Andrea Ballabio, Carlotta Giorgi

More information

RayBio Annexin V-Cy5 Apoptosis Detection Kit

RayBio Annexin V-Cy5 Apoptosis Detection Kit RayBio Annexin V-Cy5 Apoptosis Detection Kit User Manual Version 1.0 Mar 20, 2014 (Cat#: 68C5-AnnV-S) RayBiotech, Inc. We Provide You With Excellent Support And Service Tel:(Toll Free)1-888-494-8555 or

More information

Cell isolation. Spleen and lymph nodes (axillary, inguinal) were removed from mice

Cell isolation. Spleen and lymph nodes (axillary, inguinal) were removed from mice Supplementary Methods: Cell isolation. Spleen and lymph nodes (axillary, inguinal) were removed from mice and gently meshed in DMEM containing 10% FBS to prepare for single cell suspensions. CD4 + CD25

More information

Supplemental Information. Gut Microbiota Promotes Hematopoiesis to Control Bacterial Infection. Cell Host & Microbe, Volume 15

Supplemental Information. Gut Microbiota Promotes Hematopoiesis to Control Bacterial Infection. Cell Host & Microbe, Volume 15 Cell Host & Microbe, Volume 15 Supplemental Information Gut Microbiota Promotes Hematopoiesis to Control Bacterial Infection Arya Khosravi, Alberto Yáñez, Jeremy G. Price, Andrew Chow, Miriam Merad, Helen

More information

Supplementary Figure 1. Ent inhibits LPO activity in a dose- and time-dependent fashion.

Supplementary Figure 1. Ent inhibits LPO activity in a dose- and time-dependent fashion. Supplementary Information Supplementary Figure 1. Ent inhibits LPO activity in a dose- and time-dependent fashion. Various concentrations of Ent, DHBA or ABAH were pre-incubated for 10 min with LPO (50

More information

Peptide stimulation and Intracellular Cytokine Staining

Peptide stimulation and Intracellular Cytokine Staining v Peptide stimulation and Intracellular Cytokine Staining Authors A. Cosma, S. Allgayer MATERIALS Date 01-02-2007 REAGENTS: Version 1.0 - PBMCs - Culture medium - Costimulating antibodies - Peptide pools

More information

Technical Resources. BD Immunocytometry Systems. FastImmune Intracellular Cytokine Staining Procedures

Technical Resources. BD Immunocytometry Systems. FastImmune Intracellular Cytokine Staining Procedures FastImmune Intracellular Cytokine Staining Procedures BD has developed protocols for the detection of intracellular cytokines in activated lymphocytes and in activated monocytes. The procedures have been

More information

Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary Materials and Methods

Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary Materials and Methods Silva et al. PTEN posttranslational inactivation and hyperactivation of the PI3K/Akt pathway sustain primary T cell leukemia viability Supplementary Table; Supplementary Figures and legends S1-S21; Supplementary

More information

MATERIALS. Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL

MATERIALS. Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL v Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL Authors S. Kutscher, A. Cosma, MATERIALS REAGENTS: - PBMCs - Culture medium - Costimulating antibodies - Peptide pools including

More information

Figure S1. Generation of inducible PTEN deficient mice and the BMMCs (A) B6.129 Pten loxp/loxp mice were mated with B6.

Figure S1. Generation of inducible PTEN deficient mice and the BMMCs (A) B6.129 Pten loxp/loxp mice were mated with B6. Figure S1. Generation of inducible PTEN deficient mice and the BMMCs (A) B6.129 Pten loxp/loxp mice were mated with B6.129-Gt(ROSA)26Sor tm1(cre/ert2)tyj /J mice. To induce deletion of the Pten locus,

More information

PE Annexin V Apoptosis Detection Kit User Manual KT40001

PE Annexin V Apoptosis Detection Kit User Manual KT40001 PE Annexin V Apoptosis Detection Kit User Manual KT40001 For research use only. Not intended for diagnostic testing. a WuXi AppTec company www.abgent.com.cn PE Annexin-V Apoptosis Detection Kit Product

More information

For the rapid, sensitive and accurate measurement of apoptosis in various samples.

For the rapid, sensitive and accurate measurement of apoptosis in various samples. ab14082 500X Annexin V-FITC Apoptosis Detection Reagent Instructions for Use For the rapid, sensitive and accurate measurement of apoptosis in various samples. This product is for research use only and

More information

Essential Medium, containing 10% fetal bovine serum, 100 U/ml penicillin and 100 µg/ml streptomycin. Huvec were cultured in

Essential Medium, containing 10% fetal bovine serum, 100 U/ml penicillin and 100 µg/ml streptomycin. Huvec were cultured in Supplemental data Methods Cell culture media formulations A-431 and U-87 MG cells were maintained in Dulbecco s Modified Eagle s Medium. FaDu cells were cultured in Eagle's Minimum Essential Medium, containing

More information

ab Exosome Isolation and Analysis Kit - Flow Cytometry, Cell Culture (CD63 / CD81)

ab Exosome Isolation and Analysis Kit - Flow Cytometry, Cell Culture (CD63 / CD81) Version 1 Last updated 26 September 2018 ab239682 Exosome Isolation and Analysis Kit - Flow Cytometry, Cell Culture (CD63 / For the isolation and analysis of exosome from cell culture. This product is

More information

Detailed step-by-step operating procedures for NK cell and CTL degranulation assays

Detailed step-by-step operating procedures for NK cell and CTL degranulation assays Supplemental methods Detailed step-by-step operating procedures for NK cell and CTL degranulation assays Materials PBMC isolated from patients, relatives and healthy donors as control K562 cells (ATCC,

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:10.1038/nature10134 Supplementary Figure 1. Anti-inflammatory activity of sfc. a, Autoantibody immune complexes crosslink activating Fc receptors, promoting activation of macrophages, and WWW.NATURE.COM/NATURE

More information

Supporting Information

Supporting Information Supporting Information Valkenburg et al. 10.1073/pnas.1403684111 SI Materials and Methods ELISA and Microneutralization. Sera were treated with Receptor Destroying Enzyme II (RDE II, Accurate) before ELISA

More information

Influenza A H1N1 HA ELISA Pair Set

Influenza A H1N1 HA ELISA Pair Set Influenza A H1N1 HA ELISA Pair Set for H1N1 ( A/Puerto Rico/8/1934 ) HA Catalog Number : SEK11684 To achieve the best assay results, this manual must be read carefully before using this product and the

More information

Lipoprotein Lipase Activity Assay Kit (Fluorometric)

Lipoprotein Lipase Activity Assay Kit (Fluorometric) Lipoprotein Lipase Activity Assay Kit (Fluorometric) Catalog Number KA4538 100 assays Version: 02 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 General

More information

B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small cell lung cancer

B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small cell lung cancer Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2017 Experimental Methods Cell culture B16-F10 (Mus musculus skin melanoma), NCI-H460 (human non-small

More information

Human LDL Receptor / LDLR ELISA Pair Set

Human LDL Receptor / LDLR ELISA Pair Set Human LDL Receptor / LDLR ELISA Pair Set Catalog Number : SEK10231 To achieve the best assay results, this manual must be read carefully before using this product and the assay is run as summarized in

More information

Concentration Estimation from Flow Cytometry Exosome Data Protocol

Concentration Estimation from Flow Cytometry Exosome Data Protocol Concentration Estimation from Flow Cytometry Exosome Data Protocol 1. STANDARD CURVE Create a standard curve for the target exosome by plotting the mean fluorescence (y axis) against the protein concentration

More information

Annexin V-Cy3 Apoptosis Detection Reagent

Annexin V-Cy3 Apoptosis Detection Reagent ab14143 Annexin V-Cy3 Apoptosis Detection Reagent Instructions for Use For the rapid, sensitive and accurate measurement of apoptosis in various samples This product is for research use only and is not

More information

Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice

Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice (a) CD11c.DOG transgenic mice (tg) were treated with 8 ng/g body weight (b.w.) diphtheria toxin (DT) i.p. on day -1 and every

More information

Biological Consulting Services

Biological Consulting Services Biological Consulting Services of North Florida/ Inc. May 13, 2009 Aphex BioCleanse Systems, Inc. Dear Sirs, We have completed antimicrobial efficacy study on the supplied Multi-Purpose Solution. The testing

More information

SUPPLEMENTARY INFORMATION. Bacterial strains and growth conditions. Streptococcus pneumoniae strain R36A was

SUPPLEMENTARY INFORMATION. Bacterial strains and growth conditions. Streptococcus pneumoniae strain R36A was SUPPLEMENTARY INFORMATION Bacterial strains and growth conditions. Streptococcus pneumoniae strain R36A was grown in a casein-based semisynthetic medium (C+Y) supplemented with yeast extract (1 mg/ml of

More information

90 min 18 min. 45 min. 14 d

90 min 18 min. 45 min. 14 d Isolation, cultivation, and expansion of Pan T cells from human PBMCs In vitro expansion of human Pan T cells Introduction T lymphocytes (T cells) play a central role in the adaptive immune system by controlling

More information

Protocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening

Protocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening Protocol for A-549 VIM RFP (ATCC CCL-185EMT) TGFβ1 EMT Induction and Drug Screening Introduction: Vimentin (VIM) intermediate filament (IF) proteins are associated with EMT in lung cancer and its metastatic

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION Supplementary Figures Supplementary Figure S1. Binding of full-length OGT and deletion mutants to PIP strips (Echelon Biosciences). Supplementary Figure S2. Binding of the OGT (919-1036) fragments with

More information

HIV-1 p24 ELISA Pair Set Cat#: orb54951 (ELISA Manual)

HIV-1 p24 ELISA Pair Set Cat#: orb54951 (ELISA Manual) HIV-1 p24 ELISA Pair Set Cat#: orb54951 (ELISA Manual) BACKGROUND Human Immunodeficiency Virus ( HIV ) can be divided into two major types, HIV type 1 (HIV-1) and HIV type 2 (HIV-2). HIV-1 is related to

More information

Supplementary Information. Sonorensin: A new bacteriocin with potential of an anti-biofilm agent and a food

Supplementary Information. Sonorensin: A new bacteriocin with potential of an anti-biofilm agent and a food Supplementary Information Sonorensin: A new bacteriocin with potential of an anti-biofilm agent and a food biopreservative Lipsy Chopra, Gurdeep Singh, Kautilya Kumar Jena and Debendra K. Sahoo* Biochemical

More information

Supplementary Information

Supplementary Information Supplementary Information Distinct bone marrow-derived and tissue resident macrophage lineages proliferate at key stages during inflammation. 1 Luke C. Davies, 1 Marcela Rosas, 2 Stephen J. Jenkins, 1

More information

ab Annexin V- mfluor Blue 570 Detection

ab Annexin V- mfluor Blue 570 Detection Version 1 Last updated 26 March 2018 ab219914 Annexin V- mfluor Blue 570 Detection Reagent For the rapid, sensitive and accurate measurement of PS exposure in live cells This product is for research use

More information

Supplemental Figure 1. Activated splenocytes upregulate Serpina3g and Serpina3f expression.

Supplemental Figure 1. Activated splenocytes upregulate Serpina3g and Serpina3f expression. Relative Serpin expression 25 2 15 1 5 Serpina3f 1 2 3 4 5 6 8 6 4 2 Serpina3g 1 2 3 4 5 6 C57BL/6 DBA/2 Supplemental Figure 1. Activated splenocytes upregulate Serpina3g and Serpina3f expression. Splenocytes

More information

Nature Protocols: doi: /nprot Supplementary Figure 1. Fluorescent titration of probe CPDSA.

Nature Protocols: doi: /nprot Supplementary Figure 1. Fluorescent titration of probe CPDSA. Supplementary Figure 1 Fluorescent titration of probe CPDSA. Fluorescent titration of probe CPDSA (10 um) upon addition of GSH in HEPES (10 mm, ph = 7.4) containing 10% DMSO. Each spectrum was recorded

More information

Supporting Information

Supporting Information Supporting Information Identification of Novel ROS Inducers: Quinone Derivatives Tethered to Long Hydrocarbon Chains Yeonsun Hong,, Sandip Sengupta,, Wooyoung Hur, *, Taebo Sim,, * KU-KIST Graduate School

More information

Human Immunodeficiency Virus type 1 (HIV-1) p24 / Capsid Protein p24 ELISA Pair Set

Human Immunodeficiency Virus type 1 (HIV-1) p24 / Capsid Protein p24 ELISA Pair Set Human Immunodeficiency Virus type 1 (HIV-1) p24 / Capsid Protein p24 ELISA Pair Set Catalog Number : SEK11695 To achieve the best assay results, this manual must be read carefully before using this product

More information

Supporting Information

Supporting Information Supporting Information Sasportas et al. 10.1073/pnas.0806647106 SI Methods Lentiviral Transduction. MSC and glioma cells were transduced with LVs at varying multiplicity of infection (moi) by incubating

More information

ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3*

ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3* ADCC Assay Protocol Vikram Srivastava 1, Zheng Yang 1, Ivan Fan Ngai Hung 2, Jianqing Xu 3, Bojian Zheng 3 and Mei- Yun Zhang 3* 1 Department of Microbiology, Li Ka Shing Faculty of Medicine, University

More information

W/T Itgam -/- F4/80 CD115. F4/80 hi CD115 + F4/80 + CD115 +

W/T Itgam -/- F4/80 CD115. F4/80 hi CD115 + F4/80 + CD115 + F4/8 % in the peritoneal lavage 6 4 2 p=.15 n.s p=.76 CD115 F4/8 hi CD115 + F4/8 + CD115 + F4/8 hi CD115 + F4/8 + CD115 + MHCII MHCII Supplementary Figure S1. CD11b deficiency affects the cellular responses

More information

Annexin V-Cy3 Apoptosis Detection Kit

Annexin V-Cy3 Apoptosis Detection Kit ab14142 Annexin V-Cy3 Apoptosis Detection Kit Instructions for Use For the rapid, sensitive and accurate measurement of apoptosis in various samples. This product is for research use only and is not intended

More information

Luminescent platforms for monitoring changes in the solubility of amylin and huntingtin in living cells

Luminescent platforms for monitoring changes in the solubility of amylin and huntingtin in living cells Electronic Supplementary Material (ESI) for Molecular BioSystems. This journal is The Royal Society of Chemistry 2016 Contents Supporting Information Luminescent platforms for monitoring changes in the

More information

Annexin V APC Assay Kit

Annexin V APC Assay Kit Annexin V APC Assay Kit Item No. 601410 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION 3 Materials

More information

ORS 2015 Annual Meeting (Orthopedic Research Society)

ORS 2015 Annual Meeting (Orthopedic Research Society) ORS 2015 Annual Meeting (Orthopedic Research Society) Poster No: 1045 CERAMENT Bone Void Filler Impregnated with Gentamicin Increases Bone Formation and Decreases the Rate of Detectable Infection after

More information

Multi-Parameter Apoptosis Assay Kit

Multi-Parameter Apoptosis Assay Kit Multi-Parameter Apoptosis Assay Kit Catalog Number KA1335 5 x 96 assays Version: 05 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Background... 3 Principle of the Assay...

More information

Supporting Information File S2

Supporting Information File S2 Pulli et al. Measuring Myeloperoxidase Activity in Biological Samples Page 1 of 6 Supporting Information File S2 Step-by-Step Protocol Reagents: Sucrose (Sigma, S3089) CaCl 2 (Sigma, C5770) Heparin Sodium

More information

ab65336 Triglyceride Quantification Assay Kit (Colorimetric/ Fluorometric)

ab65336 Triglyceride Quantification Assay Kit (Colorimetric/ Fluorometric) Version 10 Last updated 19 December 2017 ab65336 Triglyceride Quantification Assay Kit (Colorimetric/ Fluorometric) For the measurement of triglycerides in various samples. This product is for research

More information

Data Sheet. CD28:B7-2[Biotinylated] Inhibitor Screening Assay Kit Catalog # Size: 96 reactions

Data Sheet. CD28:B7-2[Biotinylated] Inhibitor Screening Assay Kit Catalog # Size: 96 reactions Data Sheet CD28:B7-2[Biotinylated] Inhibitor Screening Assay Kit Catalog # 72062 Size: 96 reactions BACKGROUND: The activation of naïve T cells requires two signals; the specific T cell receptor recognition

More information

Supporting Information

Supporting Information Supporting Information Desnues et al. 10.1073/pnas.1314121111 SI Materials and Methods Mice. Toll-like receptor (TLR)8 / and TLR9 / mice were generated as described previously (1, 2). TLR9 / mice were

More information

Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the

Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the targeted allele in ES cells, and the mutant allele in

More information

Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic

Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic Chemical Chaperones Mitigate Experimental Asthma By Attenuating Endoplasmic Reticulum Stress Lokesh Makhija, BE, Veda Krishnan, MSc, Rakhshinda Rehman, MTech, Samarpana Chakraborty, MSc, Shuvadeep Maity,

More information

Supporting Information

Supporting Information Supporting Information lpek et al. 1.173/pnas.1121217 SI Materials and Methods Mice. cell knockout, inos / (Taconic arms), Rag1 /, INγR /, and IL-12p4 / mice (The Jackson Laboratory) were maintained and/or

More information

Supplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation

Supplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation Supplemental Materials for Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to FTY7 during neuroinflammation This file includes: Supplemental Table 1. EAE clinical parameters of

More information

RayBio Annexin V-FITC Apoptosis Detection Kit

RayBio Annexin V-FITC Apoptosis Detection Kit RayBio Annexin V-FITC Apoptosis Detection Kit User Manual Version 1.0 May 25, 2014 (Cat#: 68FT-AnnV-S) RayBiotech, Inc. We Provide You With Excellent Support And Service Tel:(Toll Free)1-888-494-8555 or

More information

Supplementalgfigureg1gSchematicgdiagramgofgtumor1modellingg

Supplementalgfigureg1gSchematicgdiagramgofgtumor1modellingg SChinjectionh F:LuchLCLsh IVhinjectionh T:cellsh Monitorhforhtumorh growthhandhxeno: reactivehgvhd GVLgexperimentg kcbgvsgpbgt1cellse Xeno1reactiveg experimentg kcbgvsgpbgt1cellse IVhinjectionh 5xh,N^6

More information

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-

(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14- 1 Supplemental Figure Legends Figure S1. Mammary tumors of ErbB2 KI mice with 14-3-3σ ablation have elevated ErbB2 transcript levels and cell proliferation (A) PCR primers (arrows) designed to distinguish

More information

O. Repeat the measurement in all relevant modes used in your experiments (e.g. settings for orbital averaging).

O. Repeat the measurement in all relevant modes used in your experiments (e.g. settings for orbital averaging). Before You Begin Read through this entire protocol sheet carefully before you start your experiment and prepare any materials you may need. This year, in order to improve reproducibility, we are requiring

More information

SHREE ET AL, SUPPLEMENTAL MATERIALS. (A) Workflow for tumor cell line derivation and orthotopic implantation.

SHREE ET AL, SUPPLEMENTAL MATERIALS. (A) Workflow for tumor cell line derivation and orthotopic implantation. SHREE ET AL, SUPPLEMENTAL MATERIALS SUPPLEMENTAL FIGURE AND TABLE LEGENDS Supplemental Figure 1. Derivation and characterization of TS1-TGL and TS2-TGL PyMT cell lines and development of an orthotopic

More information

Suppl Video: Tumor cells (green) and monocytes (white) are seeded on a confluent endothelial

Suppl Video: Tumor cells (green) and monocytes (white) are seeded on a confluent endothelial Supplementary Information Häuselmann et al. Monocyte induction of E-selectin-mediated endothelial activation releases VE-cadherin junctions to promote tumor cell extravasation in the metastasis cascade

More information

Supplementary Figure 1. Validation of astrocytes. Primary astrocytes were

Supplementary Figure 1. Validation of astrocytes. Primary astrocytes were Supplementary Figure 1. Validation of astrocytes. Primary astrocytes were separated from the glial cultures using a mild trypsinization protocol. Anti-glial fibrillary acidic protein (GFAP) immunofluorescent

More information

Influenza B Hemagglutinin / HA ELISA Pair Set

Influenza B Hemagglutinin / HA ELISA Pair Set Influenza B Hemagglutinin / HA ELISA Pair Set Catalog Number : SEK11053 To achieve the best assay results, this manual must be read carefully before using this product and the assay is run as summarized

More information