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Home › Dataset Library › Analysis of differential gene expression in Cebpa-positive and Cebpa-negative hematopoietic stem cells using a Cebpa-Cre EYFP reporter...

Dataset: Analysis of differential gene expression in Cebpa-positive and Cebpa-negative hematopoietic stem cells using a Cebpa-Cre EYFP reporter mouse model

C/EBPalpha is a transcription factor critically involved in myeloid development and indispensable for formation of granulocytes. To track...

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C/EBPalpha is a transcription factor critically involved in myeloid development and indispensable for formation of granulocytes. To track the cellular fate of stem and progenitor (LSK) cells, which express C/EBPalpha, we developed a mouse model expressing Cre recombinase from the Cebpa promoter and an inducible EYFP allele. We show that Cebpa/EYFP+ cells represent a significant subset of LSK cells, which predominantly give rise to myeloid cells in steady state hematopoiesis. C/EBPalpha induced a robust myeloid gene expression signature and downregulated E2A-induced regulators of early lymphoid development. In addition, Cebpa/EYFP+ cells comprise a fraction of early thymic progenitors (ETP) with robust myeloid potential. However, Cebpa/EYFP+ LSK and ETP cells retained the ability to develop into erythroid and T-lymphoid lineages, respectively. These findings support an instructive, but argue against a lineage restrictive role of C/EBPalpha in multipotent hematopoietic and thymic progenitors. We performed global gene expression profiling of double-sorted Cebpa/EYFP+ and Cebpa/EYFP- LSK cells of pooled Cebpa Cre/wt R26 EYFP reporter mice to identify differentially regulated genes in Cebpa+ versus Cebpa- LSK cells. RNA was isolated from three biological replicates of Cebpa/EYFP+ LSK cells and two biological replicates of Cebpa/EYFP- LSK cells. To determine if the identified genes were truly dependent on Cebpa expression, we also performed global gene expression profilling of Cebpa/EYFP+ and Cebpa/EYFP- fetal liver LSK cells of Cebpa Cre/fl R26 EYFP mice. Induction of Cebpa/Cre expression in these mice leads to Cre-mediated recombination of the floxed wt Cebpa allele resulting in a complete Cebpa knock-out. In this case, RNA was isolated from two biological replicates of either Cebpa/EYFP+ and Cebpa/EYFP- LSK cells. In addition, we included one biological replicate of Cebpa/EYFP+ and Cebpa/EYFP- fetal liver LSK cells of Cebpa Cre/wt R26 EYFP mice to determine the correlation of differentially regulated genes in bone marrow and fetal liver LSK cells.

Species:
mouse

Samples:
11

Source:
E-GEOD-23800

Updated:
Dec.12, 2014

Registered:
Nov.11, 2014


Factors: (via ArrayExpress)
Sample AGE TISSUE GENOTYPE CEBPA/EYFP INDUCTION
GSM587025 7-12 weeks bone marrow Cebpa Cre/wt R26 EYFP +
GSM587025 7-12 weeks bone marrow Cebpa Cre/wt R26 EYFP +
GSM587025 7-12 weeks bone marrow Cebpa Cre/wt R26 EYFP +
GSM587028 7-12 weeks bone marrow Cebpa Cre/wt R26 EYFP -
GSM587028 7-12 weeks bone marrow Cebpa Cre/wt R26 EYFP -
GSM587030 E14.5 fetal liver Cebpa Cre/fl R26 EYFP +
GSM587030 E14.5 fetal liver Cebpa Cre/fl R26 EYFP +
GSM587032 E14.5 fetal liver Cebpa Cre/fl R26 EYFP -
GSM587032 E14.5 fetal liver Cebpa Cre/fl R26 EYFP -
GSM587034 E14.5 fetal liver Cebpa Cre/wt R26 EYFP +
GSM587035 E14.5 fetal liver Cebpa Cre/wt R26 EYFP -

Tags

  • bone
  • bone marrow
  • liver

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