Project description:Aim of the experiment was to compare the gene expression responses of long-term hematopoietic stem cells (HSCs) and myeloid-restricted progenitors (MyPs) to the hematopoietic cell-restricted deletion of the catalytic subunit Elp3 of the Elongator complex. Since Elp3 deletion resulted in activation of the p53 pathway, we also studied the transcriptome of MyPs deficient for Trp53 or for both Elp3 and Trp53. The Elp3fl/fl strain was generated in house and first described in (DOI: 10.1084/jem.20142288). The Trp53 strain was first described in (DOI: 10.1101/gad.14.8.994) and was purchased from the Jackson Laboratory. Littermates of 8-12 weeks old were used in all experiments.
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.
Project description:In blood, the transcription factor C/EBPa is essential for myeloid differentiation and has been implicated in regulating self-renewal of fetal liver hematopoietic stem cells (HSCs). However, its function in adult HSCs is unknown. Here, using an inducible knockout model, we found that C/EBPa deficient adult HSCs underwent a pronounced expansion with enhanced proliferation, characteristics resembling fetal liver HSCs. Consistently, transcription profiling of C/EBPa deficient HSCs revealed a gene expression program similar to fetal liver HSCs. Moreover we observed that age-specific C/EBPa expression correlated with its inhibitory effect on the HSC cell cycle. Mechanistically, we identified N-Myc as a C/EBPa downstream target. C/EBPa upregulation during HSC transition from an active fetal state to a quiescent adult state was accompanied by down-regulation of N-Myc, and loss of C/EBPa resulted in de-repression of NMyc. Our data establish that C/EBPa acts as a molecular switch between fetal and adult states of HSC in part via transcriptional repression of the proto-oncogene N-Myc. HSCs of Pu.1 knock-in (PU.1ki/ki) mice were used for RNA extraction and hybridization on Affymetrix microarrays. We compared these microarray samples with the corresponding wild type.
Project description:This SuperSeries is composed of the following subset Series: GSE30444: Retroviral Sox17 over-expression adult hematopoietic stem/progenitor cells microarray GSE30445: Sox17-transgenic hematopoietic stem cell microarray Refer to individual Series
Project description:The transcriptome of Ctrl and Vitamin A-deficient longterm hematopoietic stem cells (LT-HSC) and multipotant progenitors (MPP3/4) was assessed by RNAseq.
Project description:To investigate how HSCs functionally compensate for the B cell deficiencies of other HSCs within an organism, we co-transplanted wildtype (WT) HSCs and lineage-deficient HSCs into lethally irradiated WT recipient mice. WT HSCs were then purified from the bone marrow of recipient mice for RNA isolation and sequencing. The purpose is to determine whether there are common genes shared between compensating WT HSCs in the WT co-transplanted with uMT-/- (B6.129S2(B6)-Ighmtm1Cgn/J) and WT co-transplanted with NSG (NOD-scid IL2Rgamma null).
Project description:A kinase-inactivated variant of Cdk6 (Cdk6_K43M) was associated with prolonged repopulation potential of mouse lineage-negative, Sca1-positive, cKit-positive, CD150-positive, CD48-negative (HSC/MPP1) cells. In order to understand differences between Cdk6_K43M HSC/MPP1 cells, and HSC/MPP1 cells carrying wild-type Cdk6 (Cdk6_WT) or a complete Cdk6 knock-out (Cdk6_KO) in the context of repopulation the following experiment was performed. Pools of freshly isolated CD45.2-positive HSC/MPP1 cells from mice carrying Cdk6_WT, Cdk6_K43M or Cdk6_KO (all homozygous) were serially transplanted into three CD45.1-positive recipients, per genotype. After two rounds of transplantation, CD45.2-positive HSC/MPP1 cells were isolated using flow cytometry based cell sorting and subjected to library prep for RNA sequencing.