Gene expression changes associated with kinase-inactivated Cdk6 or a complete Cdk6 KO in mouse HSC/MPP1 cells after two rounds of transplantation
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.