Bcl11b maintains quiescent mammary repopulating units
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Shang Cai
PROVIDER: E-GEOD-62059 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:This gene expression microarray analysis is to test how induced Bcl11b expression affect the overall gene expression profile of Comma D Beta Cells in the progenitor (sca1+) and differentiated (Sca1-) population. Comma D beta Cell line was cultured DMEM F12+2% FBS+1% PSA+5ug/ml insulin+10ng/ml EGF in 175 cm2 culture flask overnight, followed by 100ng/ml Doxycycline treatment for 1 day. Then, cells were trypsinized and subjected for flow sorting of Sca1+ and Sca- population. RNA was then extracted and subjected to microarray gene expression analysis.
2015-03-30 | GSE62059 | GEO
Project description:We report here on the identification and functional characterization of Sca1-positive muscle interstitial cells that contribute to regeneration or fibroadipogenic degeneration of dystrophic muscles and mediate the beneficial effects of HDAC inhibitors (HDACi) in mdx mice. We found that the phenotype adopted by these cells and their biological activity are influenced by changes in muscle environment. While Sca1-positive muscle interstitial cells from healthy muscles spontaneously adopt a fibro-adipogenic phenotype, Sca1-positive muscle interstitial cells isolated from dystrophic muscles of young mdx mice show a latent myogenic phenotype that is implemented by the exposure to HDACi. Co-culture assays in vitro and co-transplantation experiments in vivo demonstrate that HDACi also improve Sca1-positive muscle interstitial cell ability to enhance the differentiation potential of adjacent satellite cells. Importantly, HDACi-induced myogenic phenotype and pro-regeneration activity were not observed in Sca1-positive muscle interstitial cells isolated from muscles of old mdx mice. The different phenotype of Sca1-positive muscle interstitial cells from mdx mice at different stages of disease progression correlated with the stage-dependent beneficial effect of HDACi, which were effective only at early stages of disease. Transplantation of Sca1-positive muscle interstitial cells isolated from regenerating young muscles into muscles of old mdx mice restored HDACi ability to increase myofiber size. These results indicate that Sca1-positive muscle interstitial cells are key cellular determinants of disease progression and mediate the beneficial effect of HDACi in a mouse model of muscular dystrophy. Isolation of Sca1+ muscle interstitial cells from TSA treated DMD/MDX mice and MuSc from DMD/MDX mice
2012-03-31 | E-GEOD-34843 | biostudies-arrayexpress
Project description:Gene expression analyses of hematopoietic stem cells (HSCs), progenitor cells (HPCs), and differentiated cell. Gene expressions of long-term HSCs (CD34-ckit+Sca1+Lineage-), short term HSCs (CD34+ckit+Sca1+Lineage-), Progenitor cells (ckit+Sca1- Lineage-), and differentiated cels (Lineage+) were examined by microarray. Results provide insight into the mechanism of hematopoietic cell differentiation. Long-term HSCs (CD34-ckit+Sca1+Lineage-), Short term-HSCs (CD34+ckit+Sca1+Lineage-), Progenitor cells (ckit+Sca1- Lineage-, and Lineage+), and differentiated cell (Lineage+) were sorted from mouse bone marraw and were examined by microarray. Results provide insight into the mechanism of hematopoietic cell differentiation.
2013-11-06 | E-GEOD-48986 | biostudies-arrayexpress
Project description:Tomato-traced single cells from the interfollicular epidermis including the touch dome (Gli1-Tomato+/Sca1+) and the hair follicle (Gli1-Tomato+/Sca1-) were FACS-sorted, randomly sequenced and clustered into epidermal (sub-)populations.
2023-12-08 | E-MTAB-6031 | biostudies-arrayexpress
Project description:The aim of our study was to evaluate gene expression of canine mammary cancer stem-like cells co-cultured with tumor associated macrophages. Two canine mammary cancer cell lines (CMT-U27 and P114) were stained using anti-Sca1 (stem cell antigen 1), anti-EpCAM (Epithelial cell adhesion molecule) and anti-CD44 antibody. The FACS analysis showed 0,02-0,05% of Sca1+/EpCAM+/CD44+ in each of the cell line. Cancer stem-like cells were collected using FACS Aria II then co-cultured with tumor associated macrophages and used for further analysis of gene expression ( using Agilent Gene Expression Hybridization Kit ). Canine mammary cancer cell lines were stained using anti-Sca1 (stem cell antigen 1), anti-EpCAM (Epithelial cell adhesion molecule) and anti-CD44 antibodies. Next using FACS Aria II and Sca1+/EpCAM+/CD44+ cells were collected and co-cultured with tumor associated macrophages. Then, total RNA was isolated and hybridized at Gene Expression microarray.
2016-06-01 | E-GEOD-73230 | biostudies-arrayexpress
Project description:Reduced representation bisulfite sequencing (RRBS) of hematopoeitic stem/progenitor cells and mature B-cells from wild-type mice and mice transiently expressing Bcl6 under control of the Sca1 promoter. CBAxC57BL/6J mice were engineered to transiently express a Bcl6 cDNA in hematopoeitic stem/progenitor cells under control of the Sca1 promoter and then delete the transgene by cre-mediated recombination upon expression of Cd79a (Sca1-BCL6?). Hematopoeitic stem/progentior cells (HSPC, Sca1+Lin-) from the bone marrow and mature B-cells (B220+) from the peripheral blood were isolated from these mice and wild-type CBAxC57BL/6J mice by fluorescence activated cell sorting. Cells from 6-9 mice were pooled, for a total of 4 experimental conditions. DNA was extracted for each condition and analyzed by reduced representation bisulfite sequencing.
2014-03-28 | E-GEOD-56310 | biostudies-arrayexpress
Project description:Attempts at modeling chromosomal translocations involving MALT1 gene, hallmarks of human mucosa-associated lymphoid tissue (MALT) lymphoma, have failed to reproduce the disease in mice. Here we describe a transgenic model in which MALT1 expression was targeted to mouse hematopoietic stem/progenitor cells. In Sca1-MALT1 mice, MALT1 deregulation activated the NF-kappaB pathway in Sca1+ cells, promoting selective B-cell differentiation and mature lymphocyte accumulation in extranodal tissues, progressively leading to the development of clonal B-cell lymphomas. These tumors recapitulated the histopathological features of human MALT lymphomas, presenting typical lymphoepithelial lesions and plasmacytic differentiation. Transcriptional profiling of Sca1-MALT1 murine lymphomas revealed overlapping molecular signatures with human MALT lymphomas, including MALT1-mediated NFkappaB activation, pro-inflammatory signaling and XBP1-induced plasmacytic differentiation. Moreover, murine Malt1 showed proteolytic activity by cleaving Bcl10 in Sca1-MALT1 lymphomas. Our novel technological approach has allowed modeling human MALT lymphoma in mice, which represent unique tools study MALT lymphoma biology and evaluate anti-MALT1 therapies. Keywords: Genetic modification, wt vs. transgenic, MALT1 expression 8 sorting purified Sca1+Lin- samples were analized of which 5 were from Sca1-MALT1 transgenic mice and 3 were from WT mice.
2012-07-09 | E-GEOD-25636 | biostudies-arrayexpress
Project description:Attempts at modeling chromosomal translocations involving MALT1 gene, hallmarks of human mucosa-associated lymphoid tissue (MALT) lymphoma, have failed to reproduce the disease in mice. Here we describe a transgenic model in which MALT1 expression was targeted to mouse hematopoietic stem/progenitor cells. In Sca1-MALT1 mice, MALT1 deregulation activated the NF-kappaB pathway in Sca1+ cells, promoting selective B-cell differentiation and mature lymphocyte accumulation in extranodal tissues, progressively leading to the development of clonal B-cell lymphomas. These tumors recapitulated the histopathological features of human MALT lymphomas, presenting typical lymphoepithelial lesions and plasmacytic differentiation. Transcriptional profiling of Sca1-MALT1 murine lymphomas revealed overlapping molecular signatures with human MALT lymphomas, including MALT1-mediated NFkappaB activation, pro-inflammatory signaling and XBP1-induced plasmacytic differentiation. Moreover, murine Malt1 showed proteolytic activity by cleaving Bcl10 in Sca1-MALT1 lymphomas. Our novel technological approach has allowed modeling human MALT lymphoma in mice, which represent unique tools study MALT lymphoma biology and evaluate anti-MALT1 therapies. Keywords: Genetic modification, wt vs. transgenic, disease analysis, MALT lymphoma 9 samples were analized of which 5 were splenic lymphomas from Sca1-MALT1 transgenic mice and 4 were spleens from WT mice.
2012-07-09 | E-GEOD-25637 | biostudies-arrayexpress
Project description:In order to identify genes co-bound by SOX4 and SMAD3 in the context of breast cancer, different breast cell lines (HMLEs, MDA-MB-231 or HCC-1954) were used. Due to low endogenous expression levels for SOX4 in HMLEs in untreated conditions, doxycycline-dependent SOX4 overexpression was obtained by transducing HMLE cells with pIINDUCER21-SOX4 vector..Cells were plated and SOX4 and SMAD3 chromatine-immunoprecipitation was performed in untreated conditions, TGF-beta (2.5ng/ml), doxycycline (0.5ug/ml) or both as indicated. Genome-wide binding sites for SOX4 and SMAD3 was identified in HMLEs, MAD-MB-231 and HCC-1954 cells.
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Project description:Thrombopoietin (Thpo) signals via its receptor Mpl and regulates megakaryopoiesis, hematopoietic stem cell (HSC) maintenance and post-transplant expansion. Mpl expression is tightly controlled and deregulation of Thpo/Mpl-signaling is linked to hematological disorders. Here, we constructed an intracellular-truncated, signaling deficient Mpl protein which is presented on the cell surface (dnMpl). The transplantation of bone marrow cells retrovirally transduced to express dnMpl into wildtype mice induced thrombocytopenia, and a progressive loss of HSC. Functional analysis of the truncated Mpl in vitro and in vivo demonstrated no internalization after Thpo binding and the inhibition of Thpo/Mpl-signaling in wildtype cells due to dominant-negative (dn) effects by receptor competition with wildtype Mpl for Thpo binding. Gene expression analysis was performerd of Lin-Sca1+cKit+ (LSK) cells isolated from mice transplanted with dnMpl transduced BM cells. The gene expression profile supported the exhaustion of HSC due to increased cell cycle progression and identified new and known downstream effectors of Thpo/Mpl-signaling in HSC (namely TIE2, ESAM1 and EPCR detected on the HSC-enriched LSK cell population). In summary, we established a novel way of Thpo/Mpl inhibition in the adult mouse and performed in depth analysis of the phenotype including gene expression profiling. BM cells were flushed from the femurs and tibias of C57Bl/6 donor mice. Lineage-marker negative (lin-) cells were isolated by magnetic cell sorting using lineage-specific antibodies (GR1, CD11b, CD45R/B220, CD3e, TER-119). Prior to viral transduction, lin- BM cells were prestimulated for 24-48h in StemSpan, containing 10 ng/ml murine SCF, 20 ng/ml murine Thpo, 10 ng/ml recombinant human FGF-1, 20 ng/ml murine IGF2, 1% penicillin/streptomycin, 2 mM glutamine. Lin- cells were transduced twice on two following days with an MOI of 10 with retroviral vectors on Retronectin coated and preloaded wells (10 μg/cm2). On day four after isolation, 5x10^5 cells/mouse were intravenously injected into lethally irradiated C57Bl/6 mice (10 Gy). Eight weeks after transplantation Lin-Sca1+cKit+ (LSK) cells were isolated from dnMpl and control transplanted mice by fluorescent activated cell sorting. For each sample, BM of 2-5 mice were pooled. RNA was isolated using RNeasy Micro Kit (Qiagen GmbH, Hilden, Germany). RNA quality was assessed using the Agilent 2100 Bioanalyzer.
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