Mitosis. Single Nano Micro Milli Macro. Primary. PCNA expression
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1 a b c DAPI YFP CC3 DAPI YFP PCNA DAPI YFP ph3 DAPI YFP KI67 e 6 Mitosis f 1 PCNA expression %ph3 + /YFP + n= n= %PCNA+/YFP+ 8 6 Supplementary Figure 1. Proliferation/apoptosis rates of metastatic lesions. (a) Proliferation. tumor an metastases were staine for YFP (re), DAPI (blue) an Ki67 (green). (b) Mitosis. tumor an metastases were staine for YFP (re), DAPI (blue) an ph3 (green). (c) PCNA expression. tumor an metastases were staine for YFP (re), DAPI (blue) an PCNA (green). () Apoptosis. tumor an metastases were staine for YFP (re), DAPI (blue) an CC3 (green). (e) Quantification of ph3 staining; p= (f) Quantification of PCNA staining; p=.6. Bars represent means ± SD. Scale bars, 5 μm.
2 D A B C Supplementary Figure. Proliferation in human PDAC tumors an metastases. (a-c) Representative images of a micrometastatic lesion (a), gross metastasis (b) an primary tumor (c) staine for Ki67 by IHC. () Quantification of Ki67 staining. The percentage of Ki67 + cells was etermine for micrometastases (n=16), gross metastases (n=11) an primary tumors (n=7); p=.389. For large lesions (gross metastases an primary tumors), five fiels were average together. Bars represent means ± SD;, p<.1.
3 % positive cells per lesion % positive a b c ECAD ECAD ZEB1 ZEB1 % positive cells per lesion ECAD ECAD e # % positive cells per lesion # ZEB1 ZEB1 % positive cells per lesion %positive cells f ECAD ECAD # ZEB1 ZEB1 Supplementary Figure 3. Skewing of epithelial to mesenchymal cell ratio within primary tumor an metastatic lesions. (a-f) Comparison of epithelial (ECAD+, +) an mesenchymal (+, ZEB1+) cell frequency within each size category. Bars represent means ± SD;, p<.5;, p<.1; #, p<.1;, p<.1.
4 a liver met b Gross liver met c %+ area Met Gross Met e f g h %+ area Met Gross Met Supplementary Figure. Epithelial an mesenchymal features of human PDAC tumors an metastases. (a-c) Representative images of human microscopic liver metastasis (n=11), gross liver metastasis (n=) an primary PDAC tumors (n=6) staine for by IHC. () Quantification of + area per X fiel for gross metastases an primary tumors or within one cell iameter for micro-metastases. (e-g) Representative images of human microscopic liver metastasis (e), gross liver metastasis (f) an primary PDAC tumors (g) staine for by IHC. (h) Quantification of + area per X fiel for gross metastases (n=18) an primary tumors (n=6) or within one cell iameter for micro-metastases (n=57). For large lesions (gross metastases an primary tumors), five fiels were average together. Bars represent means ± SD;, p<.5;, p<.1; #, p<.1;, p<.1.
5 a Contact with myofibroblasts 1 %oflesions Number of cells 9 1 b c e DAPI YFP FN DAPI YFP SPARC DAPI YFP HABP DAPI YFP COL1 Supplementary Figure 5. Myofibroblast contact with nano-metastases epens on cell size an representative images of ECM stains. (a) Contact between nano-metastases an myofibroblasts by cell number. Data are shown as mean ± SD. P values were calculate by one sample t-tests against single cell mean;, p<.5. (b-e) Representative images of primary tumor an metastases staine for YFP (re), DAPI (blue) an (b) Collagen 1 (COL1); (c) Hyaluronic aci bining protein (HABP); () Secrete protein aciic an rich in cysteine (SPARC); an (e) Fibronectin (FN) in green. Scale bars, 5μm.
6 a liver met b Gross liver met c %FN+ area 3 1 Fibronectin Met Gross Met Supplementary Figure 6. Fibronectin eposition in human PDAC tumors an metastases. (a-c) Representative images of human microscopic liver metastasis (n=5), gross liver metastasis (n=18) an primary PDAC tumors (n=6) staine for FN by IHC. () Quantification of FN+ area per X fiel for gross metastases an primary tumors or within one cell iameter for micrometastases. For large lesions (gross metastases an primary tumors), five fiels were average together. Bars represent means ± SD;, p<.5.
7 CY liver KPCY liver a b c DAPI CD5 DAPI F/8 DAPI Gr-1 H DAPI CD3 e % positive area Leukocytes in uninvolve areas CD5 F/8 Gr-1 CD3 CY liver # KPCY liver f g h j DAPI/YFP/CD3 DAPI/YFP/Gr-1 DAPI/YFP/F/8 DAPI/YFP/CD5 % CD5+ area k l m n Leukocytes phages MDSCs %F/8+ area 8 6 %Gr-1+ area 3 1 % CD3+ area T Lymphocytes Supplementary Figure 7. Leukocytes accumulate in metastatic livers. (a-) Representative images of CD5 (a), F/8 (b), Gr-1 (c) an CD3 () staining (green) an DAPI (blue) in uninvolve areas of control (Px1-cre, Rosa Y/Y ) an metastatic (KPCY) livers. Scale bars, 5μm. (e) Quantification of leukocyte ensity in uninvolve areas of liver (CD5, p=.; F/8, p<.1; Gr-1, p<.1; CD3, p=.33). (f-j) Representative images of leukocyte ensity at metastatic lesions. Metastases an primary tumor were staine for YFP (re), DAPI (blue) an (f), CD5 (leukocytes); (g), F/8 (macrophages); (h), Gr-1 (MDSCs); an (j), CD3 (T cells) in green. Scale bars, 5μm. (k-n) Quantification of leukocyte ensity at metastatic lesions. The percent positive area for each leukocyte stain was quantifie within one cell iameter of metastatic lesions. Data are presente as mean ± SD. Data are presente as mean of the percent positive area within a X fiel ± SD. n 5 mice, 5 lesions for each stain. P values were calculate by one-way ANOVA an unpaire Stuent s t-test with Welch s correction;, p<.5;, p<.1; #, p<.1;, p<.1.
8 a Age at eath b Tumor weight 3 ays 1 grams 3 1 CTCs per ml c Circulating tumor cells DAPI YFP CC3 Supplementary Figure 8. Circulating tumor cells (CTCs) are ecrease after long-term chemotherapy. (a-b) Age at eath (a) an tumor weight (b) for untreate historical controls an treate animals. Lines represent mean ± SD. P value by Stuent s t-test. (c) CTCs in untreate (n=3) an treate (n=8) mice;, p <.5. Lines represent the mean. P value by Stuent s t- test. () Representative images of untreate an treate metastatic lesions staine for CC3 (green), YFP (re) an DAPI (blue). Scale bars, 5μm.
9 DAPI YFP ECAD a DAPI YFP c % ECAD- of YFP+ cells e ECAD- tumor cells Fsp1+ tumor cells % + of YFP+ cells b 1 6 / Liver met 8 Apoptosis after short-term treatment f 5 % CC3+ / Eca+ Eca- 3 1 Mets Supplementary Figure 9. Mesenchymal tumor cells are eplete after chemotherapy. (a) Representative images of untreate an treate (GEM/PTX for - weeks) primary tumors an small metastatic lesions staine for ECAD (green), YFP (re) an DAPI (blue). (b) Quantification of ECAD- tumor cells in untreate an treate primary tumors an small metastatic lesions. (c) Representative images of untreate an treate primary tumors an small metastatic lesions for (green), YFP (re) an DAPI (blue). () Quantification of + tumor cells in untreate an treate primary tumors an small metastatic lesions. n 7 mice for each conition. Bars represent means ± SD. Statistical significance was etermine by Stuent s t-test with Welch s correction. (e) Representative images of treate (1 ose GEM/PTX) primary tumor an liver metastasis staine for CC3 (green), ECAD (cyan), YFP (re) an DAPI (blue). Arrows enote YFP+/ECAD+/CC3+ cells, arrowheas enote YFP+/ECAD-/CC3+ cells. (f) Quantification of CC3 staining in ECAD+ an ECAD- tumor cells (n mice). Bars represent means ± SD. Statistical significance was etermine by Mann-Whitney test. Scale bars, 5 um., p<.5;, p<.1; #, p<.1;, p <.1.
10 Mouse ID Total PD1 1 1 PD1 1 1 PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD PD Supplementary Table 1. Metastatic buren (raw counts) for each animal use in Figure 1b.
11 Antiboy Concentration Company Catalog # Goat an Chicken α GFP 1:5 Abcam ab6673 Rabbit α CK19 1:1 In house N/A Rabbit α Phospho-Histone H3 1: Cell Signaling DC8 Rat α Ki67 1:1 DAKO M79 Rabbit α PCNA 1:1 Cell Signaling D3H8P Rabbit α Cleave Caspase-3 1: Cell Signaling 966 Rat α E-caherin 1:1 Invitrogen 1319 Rabbit α Clauin-7 1:1 Abcam ab787 Rabbit α Fsp1 1:5 DAKO A511 Rabbit α Zeb1 1:1 Santa Cruz sc-5388 Rabbit α αsma 1:1 Abcam ab569 Rabbit α Collagen I 1: Abcam ab371 Biotinylate bovine hyaluronic 1: Calbiochem aci bining protein Goat α SPARC 1:1 R&D Systems AF9 Rabbit α Fibronectin 1:3 Abcam ab13 Rat α CD5 1:5 BD Biosciences Rat α F/8 1:5 Ebioscience 1-81 Rat α Gr-1 1:5 Ebioscience Rat α CD3 1:5 Ebioscience 1-3 Goat α VE-caherin 1:1 R&D Systems AF1 Supplementary Table. Antiboies use for immunofluorescence an IHC.
12 Average age ± SD 61.6 ± 11.3 Average OS, mo ± SD 1.5 ± 9. Average tumor size, cm ± SD 5.6 ± 3. Sex, n (%) Male 18 (6) Female 1 () Location, n (%) Hea 18 (6) Boy 7 (3.3) Tail (13.3) Unknown, n (%) 1 (3.) Stage at iagnosis, n (%) IIB (6.7) III 7 (3.3) IV 1 (7) Surgery, n (%) (13.3) Chemotherapy, n (%) 3 (76.7) Raiation, n (%) 1 (33.3) Metastatic buren, n (%) 1 (6.7) (33.3) >1 18 (6) Differentation, n (%) Well/Moerate 1 (3.3) Moerate 1 (33.3) Moerate/Poor 1 () Poor 6 () KRAS status, n (%) Mutant 9 (96.7) WT 1 (3.3) TP53 status, n (%) Mutant 6 (86.7) WT (13.3) Supplementary Table 3. Clinicopathological characteristics of patients. Abbreviations: stanar eviation (SD); overall survival (OS); month (mo); number of patients (n); wiltype (WT).
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Supplementary Figures
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SUPPLEMENTARY FIGURES
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Supplementary Figure S1 Expression of mir-181b in EOC (A) Kaplan-Meier
Supplementary Figure S1 Expression of mir-181b in EOC (A) Kaplan-Meier curves for progression-free survival (PFS) and overall survival (OS) in a cohort of patients (N=52) with stage III primary ovarian
Supplementary Figure 1
CD31 FN Supplementary Figure 1 a Multivariate Cox regression analysis of predicting factors for disease-free and overall survival in 435 HNSCC patients b FN staining in whole sections of HNSCC c FN expression
Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at
Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at E10.5 were double-stained for TUNEL (red) and PECAM-1 (green).
AP VP DLP H&E. p-akt DLP
A B AP VP DLP H&E AP AP VP DLP p-akt wild-type prostate PTEN-null prostate Supplementary Fig. 1. Targeted deletion of PTEN in prostate epithelium resulted in HG-PIN in all three lobes. (A) The anatomy
Supplementary Figures
Supplementary Figures Supplementary Figure 1 Correlation between LKB1 and YAP expression in human lung cancer samples. (a) Representative photos showing LKB1 and YAP immunohistochemical staining in human
PHENOTYPIC DYNAMICS OF MICROGLIAL AND MONOCYTE-DERIVED CELLS IN GLIOBLASTOMA-BEARING MICE.
SUPPLEMENTARY FIGURES, TABLES AND VIDEOS PHENOTYPIC DYNAMICS OF MICROGLIAL AND MONOCYTE-DERIVED CELLS IN GLIOBLASTOMA-BEARING MICE. Clément Ricard 1,2,3,4, Aurélie Tchoghandjian 2,4, Hervé Luche 5, Pierre
Canberra, Australia). CD11c-DTR-OVA-GFP (B6.CD11c-OVA), B6.luc + and. Cancer Research Center, Germany). B6 or BALB/c.FoxP3-DTR-GFP mice were
Supplemental Materials and Methods Mice Female C57BL/6 (B6, I-E null, H-2 b ), BALB/c (H-2 d ) + ), FVB/N (H-2 q, I-E null, CD45.1 + ), and B6D2F1 (H-2 b/d ) mice were purchased from the Animal Resources
Nature Biotechnology: doi: /nbt Supplementary Figure 1
Supplementary Figure 1 Overview of the transplant procedure and supplementary data to Figure 1. a. Under isofluorane anesthesia, the lumen of the colon is washed by a gentle PBS enema. b. Using a p200
a 10 4 Link et al. Supplementary Figure 1 Nature Immunology: doi: /ni.1842 Cells per mouse ( 10 5 ) TRPV2KO anti-gr1 anti-gr anti-f4/80
a 10 4 WT 10 4 TRPV2KO 10 3 10 3 anti-gr1 10 2 10 1 anti-gr1 10 2 10 1 10 0 10 0 10 1 10 2 10 3 10 4 anti-f4/80 42.3 45.2 10 0 10 0 10 1 10 2 10 3 10 4 anti-f4/80 10 4 10 4 40 42.5 anti-cd11b 10 3 10 2
Supplementary Figure 1. Nature Neuroscience: doi: /nn.4547
Supplementary Figure 1 Characterization of the Microfetti mouse model. (a) Gating strategy for 8-color flow analysis of peripheral Ly-6C + monocytes from Microfetti mice 5-7 days after TAM treatment. Living
Supplementary Information
Supplementary Information Figure S1: Follicular melanocytes in the wound peripheral area migrate to the epidermis in response to wounding stimuli. Dorsal skin of Trp2-LacZ mice stained with X-gal and analyzed
Supplementary Table 1
Supplementary Table 1 Flow Cytometry Antibodies Antibody Fluorochrome Clone Vendor CD45 PE-cyanine 7 30-F11 D ioscience CD3 Pacific lue 17A2 iolegend (San Diego, CA) CD11b APC M1/70 iolegend (San Diego,
Evaluation of directed and random motility in microslides Assessment of leukocyte adhesion in flow chambers
Evaluation of directed and random motility in microslides Motility experiments in IBIDI microslides, image acquisition and processing were performed as described. PMN, which ended up in an angle < 180
Supplemental Figure S1. RANK expression on human lung cancer cells.
Supplemental Figure S1. RANK expression on human lung cancer cells. (A) Incidence and H-Scores of RANK expression determined from IHC in the indicated primary lung cancer subgroups. The overall expression
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
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/8/385/ra70/dc1 Supplementary Materials for The interaction of heparan sulfate proteoglycans with endothelial transglutaminase-2 limits VEGF 165 -induced angiogenesis
Supplementary Figure 1. Satellite cell contribution to myofibers in whole. gastrocnemius/plantaris/soleus, diaphragm, and EOM of 12 or 20 month
Keefe et al. p. 1 Supplementary Figure 1. Satellite cell contribution to myofibers in whole muscles. (a-l) Representative cross-sections through whole TA/EDL, gastrocnemius/plantaris/soleus, diaphragm,
Multiple mechanisms underlying acquired resistance to taxanes in selected docetaxelresistant MCF-7 breast cancer cells
Wang et al. BMC Cancer 2014, 14:37 RESEARCH ARTICLE Multiple mechanisms underlying acquired resistance to taxanes in selected docetaxelresistant MCF-7 breast cancer cells Harris Wang 1, The Vo 1, Ali Hajar
MATERIALS AND METHODS. Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All
MATERIALS AND METHODS Antibodies (Abs), flow cytometry analysis and cell lines Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All other antibodies used
* * * * Supplementary Figure 1. DS Lv CK HSA CK HSA. CK Col-3. CK Col-3. See overleaf for figure legend. Cancer cells
Supplementary Figure 1 Cancer cells Desmoplastic stroma Hepatocytes Pre-existing sinusoidal blood vessel New blood vessel a Normal liver b Desmoplastic HGP c Pushing HGP d Replacement HGP e f g h i DS
SUPPLEMENTARY FIG. S2. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of dnscs.
Supplementary Data SUPPLEMENTARY FIG. S1. Representative counting fields used in quantification of the in vitro neural differentiation of pattern of anpcs. A panel of lineage-specific markers were used
Supplemental Information. Tissue-Resident Macrophages in Pancreatic. Ductal Adenocarcinoma Originate from Embryonic
Immunity, Volume 7 Supplemental Information Tissue-Resident Macrophages in Pancreatic Ductal Adenocarcinoma Originate from Embryonic Hematopoiesis and Promote Tumor Progression Yu Zhu, John M. Herndon,
Supplemental Figures:
Supplemental Figures: Figure 1: Intracellular distribution of VWF by electron microscopy in human endothelial cells. a) Immunogold labeling of LC3 demonstrating an LC3-positive autophagosome (white arrow)
Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD-
Supplementary Methods Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- L1 (10F.9G2, rat IgG2b, k), and PD-L2 (3.2, mouse IgG1) have been described (24). Anti-CTLA-4 (clone
Nature Medicine: doi: /nm.4078
Supplementary Figure 1. Cetuximab induces ER stress response in DiFi cells. (a) Scheme of SILAC proteome. (b) MS-base read out of SILAC experiment. The histogram of log 2 -transformed normalized H/L ratios
Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence
Supplemental Table 1. Primers used for RT-PCR analysis of inflammatory cytokines Gene Primer Sequence IL-1α Forward primer 5 -CAAGATGGCCAAAGTTCGTGAC-3' Reverse primer 5 -GTCTCATGAAGTGAGCCATAGC-3 IL-1β
SSM signature genes are highly expressed in residual scar tissues after preoperative radiotherapy of rectal cancer.
Supplementary Figure 1 SSM signature genes are highly expressed in residual scar tissues after preoperative radiotherapy of rectal cancer. Scatter plots comparing expression profiles of matched pretreatment
Supplementary Figure 1: STAT3 suppresses Kras-induced lung tumorigenesis
Supplementary Figure 1: STAT3 suppresses Kras-induced lung tumorigenesis (a) Immunohistochemical (IHC) analysis of tyrosine 705 phosphorylation status of STAT3 (P- STAT3) in tumors and stroma (all-time
Supplementary Figure 1: Fn14 is upregulated in the epidermis and dermis of mice
Supplementary Figure 1: Fn14 is upregulated in the epidermis and dermis of mice undergoing AD- and psoriasis-like disease. Immunofluorescence staining for Fn14 (green) and DAPI (blue) in skin of naïve
Supplementary Figure S1. PTPN2 levels are not altered in proliferating CD8+ T cells. Lymph node (LN) CD8+ T cells from C57BL/6 mice were stained with
Supplementary Figure S1. PTPN2 levels are not altered in proliferating CD8+ T cells. Lymph node (LN) CD8+ T cells from C57BL/6 mice were stained with CFSE and stimulated with plate-bound α-cd3ε (10µg/ml)
(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