a b Pml F/F Prrx1-cre f 1.4 n.s.
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1 a b Pml F/F Prrx1-re X Prrx1-re-Pml F/F Relative CFU-F numbers 2. Prrx1-Cre-PML +/+ n.s. d early passages e f 1.4 n.s. Adsorbane Adsorbane days in ulture late passages days in ulture Osteoblasts ALP staining Pml +/+ Pml -/- Relative expression Pml mrna BM-MSCs MSCs CD51+Sa1+ Stroma CD51-Sa1- HSCs Prrx1-Cre Pml +/+ Prrx1-Cre Pml F/F Adipoytes Oil-Red-O staining Pml +/+ Pml -/- Prrx1-Cre Pml +/+ Prrx1-Cre Pml F/F Relative adipoyti olonies/ total olonies Prrx1-Cre Pml +/+ Prrx1-Cre Pml F/F Relative expression Fabp4 mrna Prrx1-Cre-PML +/+ A. Prrx1-Cre-PML +/+ Relative expression Alp mrna A. Prrx1-Cre-PML +/+ O. Prrx1-Cre-PML +/+ O. Supplementary Figure 1 Pml regulates the biology of MSCs. (a) Shemati overview of the experimental design used to generate Pml-floxed mie. (b) Expression levels, assayed by RT-qPCR, of Pml mrna in the different populations of mesenhymal ells of Prrx1-Cre-Pml +/+ or Prrx1-Cre-Pml F/F mie; HSCs are used as ontrol. Cells olleted from n=3 mie for eah genotype were pooled together prior to RNA extration. () CFU-F olonies forming the apaity of MSCs derived from Prrx1-Cre- Pml F/F or Prrx1-Cre-Pml +/+ mie (n=7). (d) Proliferation of MSCs derived from Pml +/+ or Pml -/- mie either at early passages in vitro, or at late passages. One representative experiment is shown. (e) Differentiation of MSCs derived from Pml +/+ or Pml -/- mie into osteoblasts or adipoytes. Osteoblasts (on the left) were stained with Alkalyne Phosphatase (ALP), while adipoytes (on the right) were stained Oil-Red-O. The hart on the right shows the quantifiation of Oil-Red-O positive olonies (n=4). (f) Capaity of Prrx1-Cre-Pml +/+ or Prrx1-Cre-Pml F/F MSCs to differentiate to adipoytes (upper hart, A= MSCs triggered to adipogenesis), or into osteoblasts (lower hart. O= MSCs triggered to osteogenesis) one triggered with speifi fators. The hart on the upper right panel shows the relative expression of Fabp4, while the one on the bottom shows the expression of Alp. One representative experiment ± SEM is shown.
2 a CD48 Gated on CD34- KLS ells Gated on KLS ells Prrx1-Cre-Pml +/+ Prrx1-Cre-Pml F/F Prrx1-Cre-Pml +/+ Prrx1-Cre-Pml F/F C-kit CD15 CD34 Gated on Lin- ells Prrx1-Cre-Pml +/+ Prrx1-Cre-Pml F/F CD34- KLS ells 2. Prrx1-Cre n.s. Prrx1-Cre PmlF/F C-kit Sa1 b BA+ + leukemi ells 1 st Reipients 2 nd Reipients Endosteal BM (E-BM) Pml +/+ Pml -/-.9%.4% Inner BM (I-BM) Pml +/+ Pml -/- 21.7% 8.5% numbers of BA++KLS ells numbers of BA++KLS ells 1 st Reipients 2 nd Reipients % survival % survival BCR/ABL leukemi ells days BCR/ABL leukemi ells days Control Control Supplementary Figure 2 Pml expressed in MSCs regulates only marginally non-ellautonomously HSCs. (a) Representative plots showing the hematopoieti sub-populations in Prrx1- Cre-Pml +/+ or Prrx1-Cre-Pml F/F mie. The upper left plot shows CD15+CD48-CD34-KLS ells; the one on the right shows CD34-KLS ells, while the one on the bottom shows KLS ells. The hart on the right shows the quantifiation of CD34-KLS ells in Prrx1-Cre-Pml +/+ or Prrx1-Cre-Pml F/F mie (n=5 mie analyzed). (b) Relative perentages (representative plots on the left) and relative total numbers (harts on the right) of BA + + KLS ells present at the endosteal bone marrow (E-BM), or within the inner avity of the bone marrow (I-BM) in seond reipients Pml +/+ or Pml -/- (n=2 mie analyzed). () Survival urves of serially transplanted Prrx1-Cre-Pml F/F or ontrol mie with BA+ + ells. The upper panel shows the survival of primary reipients, while the bottom panel shows the survival of seondary reipient mie.
3 a ~ 1 passages Day 1% oxygen Day 12 leukemi ells 1, 1, Total leukemi ells and LICs Sorting MSCs Freshly isolated MSCs b wild type SCL-tTA/BCR-ABL gated on Lin- ells KLS ells RV- Fusion gene DNA Pre-leukemi ells + HoxA9 Meis- 1 MLL- AF9 Expansion in wild type reipients + leukemi ells SERIAL o-ultures with MSCs ( or ) BA+KLS ells LV- numbers of BA++ ells d 1 st o-ulture HoxA9-Meis1++ ells 9.8% 6.1% MLL-AF9-+ ells numbers of BA++KLS ells +kit+ ells st o-ulture e 3 rd o-ulture 12% 4.9% Br/Abl HoxA9+ Meis1 +kit+ ells %.6%
4 Supplementary Figure 3 Pml expressed in MSCs regulates non-ell-autonomously LICs. (a) Shemati overview of MSCs ulturing onditions, and of the o-ultures with MSCs and leukemi ells. (b) Shemati overview of the experimental design to generate leukemi ells that express HoxA9-Meis1,, or +BCR/ABL. () BA++ ells, HoxA9+Meis1 leukemi ells and leukemi ells derived from the first o-ultures with MSCs Pml +/+ or Pml -/- (n 4). (d) Relative numbers and perentages of BA++KLS ells, HoxA9- Meis1 ++ ells, and ++ ells in the first o-ultures with Pml +/+ or Pml -/- MSCs. Quantifiations are represented in the harts on the right, while representative plots are shown on the left (n 3). (e) Representative plots showing perentages of BA++KLS ells, HoxA9-Meis1 ++ ells, and ++ ells in serial o-ultures with Pml +/+ or Pml -/- MSCs.
5 b From o-ultures with MSCs Pml +/+ a Relative number of olonies HoxA9-Meis1 Relative number of olonies From o-ultures with MSCs Pml -/- % survival 15 PML+/+ PML-/- 1 5 d 1x 4x From o-ultures with MSCs Pml +/+ 1x Untreated AS 2 O 3 6x 1x From o-ultures with MSCs Pml -/- 1x Pml Pml Β-atin shscr shpml Β-atin e Relative number of olonies % survival Methyl ellulose Untreated AS 2 O 3 shscr shpml days 1ells PML+/+ 1ells PML-/- 2ells PML+/+ 2ells PML-/ days
6 Supplementary Figure 4 Pml expressed in MSCs regulates leukemi ells in a non-ellautonomous manner. (a) Relative number of olonies in methyl-ellulose, generated by leukemi ells derived from o-ultures with MSCs Pml +/+ or Pml -/-. HoxA9-Meis1 leukemi ells are shown on the left, while leukemi ells are shown on the right. One experiment ± SEM is shown. (b) H&E of leukemi blasts in the blood of mie transplanted with HoxA9-Meis1 leukemi ells after o-ulture with or MSCs. The survival urve of these mie is shown on the right. () H&E of leukemi blasts in the blood of mie transplanted with leukemi ells after oulture with or MSCs. The survival urve of these mie is shown on the right. (d) Western blot analysis of Pml expression in MSCs upon knok-down with shrna and upon treatment with AS 2 O 3. (e) Methyl-ellulose assay performed with leukemi ells () whih have been o-ultured with MSCs, while treating with AS 2 O 3, or performed with leukemi ells o-ultured with MSCs transdued with an shrna targeting Pml. One experiment ± SEM is shown.
7 a 1 st o-ulture 2 nd o-ulture BCR/ABL H+M Start Treatment 1 st o-ulture d Treatment: Anti-CXCR2 Anti-IL6R End 1 st o-ulture: LICs Start 2 nd o-ulture BCR/ABL H+M Day Day 1 Day 4 Day 8 f +Sa1+kit+ ells +kit+ ells Anti-CXCR2 Anti-IL6R Analysis of LICs N.T. HoxA9+Meis1 N.T. b acxcr2 BCR/ABL BCR/ABL ail6r HoxA9+Meis1 15% 9.89% 1.2% 6.9% acxcr2+ail6r 16% 9% 1 st o-ulture 2 nd o-ulture + + Not Treated (N.T.) CXCL1+IL6 reombinant Total numbers of leukemi ells x1 3 4 e kit+ ells MSCs Pml +/+ N.T MSCs Pml +/+ acxcr2+ail6r + N.T. 12% MSCs Pml -/- N.T 15.4% 7.9% 6.2% Total numbers of leukemi ells x MSCs Pml -/- CXCL1+IL % acxcr2+ail6r acxcr2+ail6r % of +kit+ ells Total numbers of leukemi ells x % Pml +/+ Pml -/- Pml +/+ acxcr2+ail6r Pml -/- CXCL1+IL6 +
8 Supplementary Figure 5 Pml regulates leukemi ells non-ell-autonomously through IL6/IL6R and CXCL1/CXCR2 pathways. (a) Shemati overview of the experimental design for treatment of the o-ultures with anti-il6r and anti-cxcr2 antibodies. (b) Total number of leukemi ells not treated () or treated with anti-il6r, anti-cxcr2 singularly or in ombination. One representative experiment ± SEM is shown. () Total numbers of BA++KLS ells after the treatment with anti- IL6R or/and anti-cxcr2, singularly or in ombination. Not treated ells () were used as ontrol. The hart on the left shows the quantifiation, while representative plots are shown on the right. One representative experiment ± SEM is shown. (d) Chart on the left shows the numbers of HoxA9- Meis1++kit+ ells after the first o-ulture and treatment with anti-il6r or/and anti-cxcr2 antibodies (singularly or in ombination) One representative experiment ± SEM is shown; representative plots are shown on the right. (e) Chart on the left shows the numbers of MLL-AF9+ +kit+ ells after the first o-ulture and treatment with anti-il6r or/and anti-cxcr2 antibodies (singularly or in ombination) One representative experiment ± SEM is shown; representative plots are shown on the right. (f) Shemati overview of the experimental design is shown on the left. Coultures of MSCs Pml +/+ and HoxA9-Meis1 + + leukemi ells were untreated () as ontrol, or treated with anti-cxcr2 in ombination of anti-il6r. Co-ultures with MSCs Pml -/- and HoxA9- Meis1 + + leukemi ells were untreated as ontrol, or treated with reombinant Il6 and Cxl1 proteins. Leukemi ells were then re-plated onto the new MSCs (Pml +/+ or Pml -/- ); seondary oultures were analyzed. Perentages of +kit+ ells in the different onditions are shown in the plots and the hart on the right. One experiment ± SEM is shown.
9 Supplementary Figure 6 Original blots of Figure 5
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Supplementary Figure 1 Differential expression of mirnas from the pri-mir-17-92a locus. (a) The mir-17-92a expression unit in the third intron of the host mir-17hg transcript. (b,c) Impact of knockdown
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SUPPLEMENTARY MATERIAL IL-1 signaling modulates activation of STAT transcription factors to antagonize retinoic acid signaling and control the T H 17 cell it reg cell balance Rajatava Basu 1,5, Sarah K.
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DOI: 1.138/ncb3355 a S1A8 + cells/ total.1.8.6.4.2 b S1A8/?-Actin c % T-cell proliferation 3 25 2 15 1 5 T cells Supplementary Figure 1 Inter-tumoral heterogeneity of MDSC accumulation in mammary tumor
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SUPPLEMENTARY INFORMATION doi:.8/nture89 4 4 Ilr -/- Ilr -/- Ilr -/- Cspse- -/- As -/- Nlrp -/- Il8 -/- Ilr -/- Supplementl figure. Inresed severity of NASH in inflmmsome-defiient mie, ut not in Ilr-defiient
Supplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS)
Supplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS) and their exosomes (EXO) in resting (REST) and activated
sirna count per 50 kb small RNAs matching the direct strand Repeat length (bp) per 50 kb repeats in the chromosome
Qi et al. 26-3-2564C Qi et al., Figure S1 sirna count per 5 kb small RNAs matching the direct strand sirna count per 5 kb small RNAs matching the complementary strand Repeat length (bp) per 5 kb repeats
% of Nestin-EGFP (+) cells
8 7 6 3 Nestin CD % of Nestin-EGFP (+) ells 8 6 Nestin-EGFP Marge Nestin-EGFP Marge Expression levels of Expression levels of Tumorigeniity y stemness markers ifferentiation markers limiting ilution assay
Hematopoiesis. - Process of generation of mature blood cells. - Daily turnover of blood cells (70 kg human)
Hematopoiesis - Process of generation of mature blood cells - Daily turnover of blood cells (70 kg human) 1,000,000,000,000 total cells 200,000,000,000 red blood cells 70,000,000,000 neutrophils Hematopoiesis
Quantitative PPARγ expression affects the balance between tolerance and immunity
Quantitative PPARγ expression affects the balance between tolerance and immunity Ya-Hui Liu 1, Yau-Sheng Tsai 1,2,3, Shih-Chieh Lin 4, Nan-Shih Liao 5, Ming-Shiou Jan 6, Chung-Tiang Liang 7, Shih-Wen Hsu
Supplementary Figure 1
Supplementary Figure 1 A B mir-141, human cell lines mir-2c, human cell lines mir-141, hepatocytes mir-2c, hepatocytes Relative RNA.1.8.6.4.2 Relative RNA.3.2.1 Relative RNA 1.5 1..5 Relative RNA 2. 1.5
Transduction of lentivirus to human primary CD4+ T cells
Transduction of lentivirus to human primary CD4 + T cells Human primary CD4 T cells were stimulated with anti-cd3/cd28 antibodies (10 µl/2 5 10^6 cells of Dynabeads CD3/CD28 T cell expander, Invitrogen)
pro-b large pre-b small pre-b CCCP (µm) Rag1 -/- ;33.C9HCki
a TMRM FI (Median) b TMRM FI (Median) c 20 15 10 5 0 8 6 4 2 0 pro-b large pre-b small pre-b 0 10 20 30 40 50 60 70 80 90 100 TMRM (nm) pro-b large pre-b small pre-b 0 1 2 4 8 16 32 64 128 256 CCCP (mm)
Supplementary Data. (continued)
Supplementary Data SUPPLEMENTARY FIG. S1. Gating strategy for Figure 1. Splenocytes were gated for dimension and for positivity to the different markers representing different cell populations. (A) B cells,
Akt and mtor pathways differentially regulate the development of natural and inducible. T H 17 cells
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Soft Agar Assay. For each cell pool, 100,000 cells were resuspended in 0.35% (w/v)
SUPPLEMENTARY MATERIAL AND METHODS Soft Agar Assay. For each cell pool, 100,000 cells were resuspended in 0.35% (w/v) top agar (LONZA, SeaKem LE Agarose cat.5004) and plated onto 0.5% (w/v) basal agar.
Supporting Information Table of Contents
Supporting Information Table of Contents Supporting Information Figure 1 Page 2 Supporting Information Figure 2 Page 4 Supporting Information Figure 3 Page 5 Supporting Information Figure 4 Page 6 Supporting
Nature Medicine: doi: /nm.3922
Title: Glucocorticoid-induced tumor necrosis factor receptor-related protein co-stimulation facilitates tumor regression by inducing IL-9-producing helper T cells Authors: Il-Kyu Kim, Byung-Seok Kim, Choong-Hyun
SD-1 SD-1: Cathepsin B levels in TNF treated hch
SD-1 SD-1: Cathepsin B levels in TNF treated hch. A. RNA and B. protein extracts from TNF treated and untreated human chondrocytes (hch) were analyzed via qpcr (left) and immunoblot analyses (right) for
Control shrna#9 shrna#12. shrna#12 CD14-PE CD14-PE
a Control shrna#9 shrna#12 c Control shrna#9 shrna#12 e Control shrna#9 shrna#12 h 14 12 CFU-E BFU-E GEMM GM b Colony number 7 6 5 4 3 2 1 6 pm A pa pc CFU-E BFU-E GEMM GM pu pgm A p pg B d f CD11b-APC
Supplementary. presence of the. (c) mrna expression. Error. in naive or
Figure 1. (a) Naive CD4 + T cells were activated in the presence of the indicated cytokines for 3 days. Enpp2 mrna expression was measured by qrt-pcrhr, infected with (b, c) Naive CD4 + T cells were activated
Nature Genetics: doi: /ng Supplementary Figure 1. HOX fusions enhance self-renewal capacity.
Supplementary Figure 1 HOX fusions enhance self-renewal capacity. Mouse bone marrow was transduced with a retrovirus carrying one of three HOX fusion genes or the empty mcherry reporter construct as described
Supplementary Materials
Supplementary Materials Figure S1. MTT Cell viability assay. To measure the cytotoxic potential of the oxidative treatment, the MTT [3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide] assay
Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk
Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk -/- mice were stained for expression of CD4 and CD8.
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DOI:.38/ncb3399 a b c d FSP DAPI 5mm mm 5mm 5mm e Correspond to melanoma in-situ Figure a DCT FSP- f MITF mm mm MlanaA melanoma in-situ DCT 5mm FSP- mm mm mm mm mm g melanoma in-situ MITF MlanaA mm mm
Inhibition of TGFβ enhances chemotherapy action against triple negative breast cancer by abrogation of
SUPPLEMENTAL DATA Inhibition of TGFβ enhances chemotherapy action against triple negative breast cancer by abrogation of cancer stem cells and interleukin-8 Neil E. Bhola 1, Justin M. Balko 1, Teresa C.
Zhu et al, page 1. Supplementary Figures
Zhu et al, page 1 Supplementary Figures Supplementary Figure 1: Visual behavior and avoidance behavioral response in EPM trials. (a) Measures of visual behavior that performed the light avoidance behavior
Nature Immunology: doi: /ni Supplementary Figure 1
Supplementary Figure 1 NLRP12 is downregulated in biopsy samples from patients with active ulcerative colitis (UC). (a-g) NLRP12 expression in 7 UC mrna profiling studies deposited in NCBI GEO database.
Supplemental Figure 1. (A) Western blot for the expression of RIPK1 in HK-2 cells treated with or without LPS (1 µg/ml) for indicated times.
Supplemental Figure 1. (A) Western blot for the expression of RIPK1 in HK-2 cells treated with or without LPS (1 µg/ml) for indicated times. Western blots shown are representative results from 3 independent
<10. IL-1β IL-6 TNF + _ TGF-β + IL-23
3 ns 25 ns 2 IL-17 (pg/ml) 15 1 ns ns 5 IL-1β IL-6 TNF
Cytomegalovirus and tumor stress-surveillance by human γδ T cell receptor binding to Endothelial Protein C Receptor
Cytomegalovirus and tumor stress-surveillane y human γδ T ell reeptor inding to Endothelial Protein C Reeptor Carrie R. Willox, Vinent Pitard, Sonia Netzer, Lionel Couzi, Mahoo Salim, Toias Silerzahn,
Pearson r = P (one-tailed) = n = 9
8F4-Specific Lysis, % 1 UPN1 UPN3 8 UPN7 6 Pearson r =.69 UPN2 UPN5 P (one-tailed) =.192 4 UPN8 n = 9 2 UPN9 UPN4 UPN6 5 1 15 2 25 8 8F4, % Max MFI Supplementary Figure S1. AML samples UPN1-UPN9 show variable
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/2/1/ra81/dc1 Supplementary Materials for Delivery of MicroRNA-126 by Apoptotic Bodies Induces CXCL12- Dependent Vascular Protection Alma Zernecke,* Kiril Bidzhekov,
Nature Immunology: doi: /ni Supplementary Figure 1. Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice.
Supplementary Figure 1 Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice. (a) Gene expression profile in the resting CD4 + T cells were analyzed by an Affymetrix microarray
Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured
Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured under Th0, Th1, Th2, Th17, and Treg conditions. mrna
Supplementary Figure 1. mtor LysM and Rictor LysM mice have normal cellularity and percentages of hematopoe>c cells. a. Cell numbers of lung, liver,
a. b. c. Supplementary Figure 1. mtor LysM and Rictor LysM mice have normal cellularity and percentages of hematopoe>c cells. a. Cell numbers of lung, liver, and spleen. b. Cell numbers of bone marrow
ILC1 and ILC3 isolation and culture Following cell sorting, we confirmed that the recovered cells belonged to the ILC1, ILC2 and
Supplementary Methods and isolation and culture Following cell sorting, we confirmed that the recovered cells belonged to the, ILC2 and subsets. For this purpose we performed intracellular flow cytometry
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doi: 1.138/nature89 IFN- (ng ml ) 5 4 3 1 Splenocytes NS IFN- (ng ml ) 6 4 Lymph node cells NS Nfkbiz / Nfkbiz / Nfkbiz / Nfkbiz / IL- (ng ml ) 3 1 Splenocytes IL- (ng ml ) 1 8 6 4 *** ** Lymph node cells
Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was
Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was sorted by FACS. Surface markers for sorting were CD11c +
Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4
Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4 Ankita Garg, Rodney Trout and Stephen A. Spector,,* Department
CD4 + T cells recovered in Rag2 / recipient ( 10 5 ) Heart Lung Pancreas
a CD4 + T cells recovered in Rag2 / recipient ( 1 5 ) Heart Lung Pancreas.5 1 2 4 6 2 4 6 Ctla4 +/+ Ctla4 / Ctla4 / Lung Ctla4 / Pancreas b Heart Lung Pancreas Ctla4 +/+ Ctla4 / Ctla4 / Lung Ctla4 / Pancreas