Hdac3 is essential for DNA replication and differentiation of hematopoietic stem and progenitor cells

ABSTRACT: Background: To define changes in gene expression from stem cells and early progenitor cells lacking histone deacetylase 3 (Hdac3), we purified bone marrow Lineage Negative, Sca1/cKit positive and Flt3 negative cells from wild type and Vav-Cre/Hdac3Flox/- mice. These lineage-specific knock out mice lack Hdac3 throughout the hematopoietic system. To ensure that only cells lacking Hdac3 were measured, we used a Lox-STOP-Lox-ROSA26-GFP transgene such that any cell containing active Cre also expresses GFP. Methods: Bone marrow cells were harvested from 10-30 mice and the lineage negative fraction was separated using the Lineage Cell Depletion Kit and MACS columns (Miltenyi Biotec). The lineage negative fraction was then stained with antibodies for flow cytometry and the GFP positive fraction of the LSK/Flt3 cells were sorted on a Becton Dickinson FACSAria. Total RNA was isolated from the sorted bone marrow cells using a PerfectPure RNA extraction kit (5 Prime). LSK/Flt3- cells pooled from 2 groups of 5 null mice were compared to LSK/Flt3- or LSK/Flt3+ cells pooled from 30 wild type mice. The expression of individual genes was verified using reverse transcriptase (RT) PCR. Conclusion: Hematopoietic stem and early progenitor cells fail to express gene that are typically turned on early during lymphoid development.

ORGANISM(S): Mus musculus

PROVIDER: GSE45952 | GEO | 2015/01/09

SECONDARY ACCESSION(S): PRJNA196712

REPOSITORIES: GEO