Project description:To determine the molecular mechanisms involved in the compromised erythropoiesis of ESAM-KO mice, Lin- FCgRII/III-/Lo CD41Lo c-Kit+ Sca1- endoglin+ CD150+ cells, which contain mostly BFU-E and CFU-E, were sorted from 5-FU-treated WT and ESAM-KO mice, and were subjected to microarray analyses.
Project description:This SuperSeries is composed of the following subset Series: GSE37028: Microarray analysis of Zbtb46 KO CD4+ Splenic DCs and bone marrow erythroid progenitors GSE37029: Microarray analysis of WT bone marrow myeloid progenitors, BM cultured with GM-CSF and M-CSF, and monocytes treated with GM-CSF Refer to individual Series
Project description:LSKs(lineage-, Sca-1+, c-kit+ cells) in the bone marrow from normal and 5-Fluorouracil (5-FU) treated (Day 9) wild type (WT) and Tespa1-knockout (KO) mice were isolated to explore the differentially expressed genes by RNA-seq.
Project description:Dendritic cells (DCs) in tissues and lymphoid organs comprise distinct functional subsets that differentiate in situ from circulating progenitors. Tissue-specific signals that regulate DC subset differentiation are poorly understood. We report that DC-specific deletion of the Notch2 receptor caused a reduction of DC populations in the spleen. Within the splenic CD11b+ DCs, Notch signaling blockade ablated a distinct population marked by high expression of adhesion molecule Esam. The Notch-dependent Esamhi DC subset also required lymphotoxin beta receptor signaling, proliferated in situ and facilitated efficient CD4+ T cell priming. The Notch-independent Esamlo DCs expressed monocyte-related genes and showed superior cytokine responses. In addition, Notch2 deletion led to the loss of CD11b+ CD103+ DCs in the intestinal lamina propria and to the corresponding decrease of IL-17-producing CD4+ T cells in the intestine. Thus,Notch2 is a common differentiation signal for T cell-priming CD11b+ DC subsets in the spleen and intestine. We compared genome-wide expression profiles of wild-type Esam(hi) and Esam(lo) splenic CD11b+ DC populations, along with CD11b+ DCs from DC-RBPJΔ mice. Spleens from 2-3 Cx3cr1-GFP+ RBPJflox/flox CD11c-Cre+ mice or Cx3cr1-GFP+ RBPJflox/flox Cre-negative littermate controls were isolated, pooled and depleted of lymphoid and erythroid cells by negative selection on MACS columns. Live cells were stained for surface expression of CD11c, CD11b and Esam. CD11c(hi) CD11b+ DCs from control mice could be separated into Esam(lo) GFP(hi) versus Esam(hi) GFP(lo) subsets. CD11c(hi) CD11b+ DCs from RBPJ-targeted mice spleens were uniformly Esam(lo) GFP(hi). The two subsets from control mice and single Esam(lo) GFP(hi) subset from RBPJ-targeted mice were sorted using FACSAria II flow sorter and analyzed using GeneChip Mouse Gene 1.0 ST Array (Affymetrix).
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare TP53-WT and TP53-KO HCT116 cells transcriptome profiling (RNA-seq) under 5-FU treatment condition and to evaluate the correlation between transcriptome profileing and chromatin accessibility under 5-FU treatment. Methods: HCT116 cell profiles of TP53-WT and TP53- KO were generated by deep sequencing, in duplicates, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform levels: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA). Conclusions: Our study represents the detailed analysis of TP53-WT and TP53-KO HCT116 cell transcriptomes under 5-FU treatment with different timepoint, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that RNA-seq offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a TP53-WT and TP53-KO cells with and without 5-FU treatment in different timepoint. We conclude that NGS based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.
Project description:We previously identified endothelial cell-selective adhesion molecule (ESAM) as a novel functional marker of hematopoietic stem cells (HSCs) in mice. To examine ESAM expression in human, we analyzed diverse HSC sources using flow cytometry. From mononuclear cells of collagenase-treated human hip bones, we obtained two ESAM positive populations in CD34(+) CD38(-) fraction, referred to as ESAM(High) and ESAM(Bright). Then we conducted microarray analyses comparing gene expression signatures between these two populations.
Project description:We previously identified endothelial cell-selective adhesion molecule (ESAM) as a novel functional marker of hematopoietic stem cells (HSCs) in mice. To examine ESAM expression in human, we analyzed diverse HSC sources using flow cytometry. From mononuclear cells of collagenase-treated human hip bones, we obtained two ESAM positive populations in CD34(+) CD38(-) fraction, referred to as ESAM(High) and ESAM(Bright). Then we conducted microarray analyses comparing gene expression signatures between these two populations. Trabecular tissues of the femora were treated with collagenase IV and DNase. Mononuclear cells were collected, and CD34(+) CD38(-) ESAM(High) or ESAM(Bright) cells were sorted.