Genes regulated by HEB and E2A during DP stage of thymocyte development. Mus musculus

To investigate gene targets of the E-proteins HEB and E2A during the CD4+CD8+ double positive (DP) stage of T cell development. We examined E-protein function by simultaneous removal of both HEB (Tcf12) and E2A (Tcfe2a) genes at the DP stage. This was done by crossing mice containing HEB floxed and E2A floxed alleles to a CD4Cre background (Tcf12f/fTcfe2af/fCD4Cre mice). Microarray analysis was used to compare gene expression in HEB and E2A double deficient DP thymocytes (Cre+) to Cre- control DP thymocytes. Keywords: genetic modification Overall design: CD4+CD8+ DP cells were sorted from Tcf12f/fTcfe2af/fCD4Cre+ (Cre+) and Tcf12f/fTcfe2af/fCD4Cre- (Cre-) thymus. The same CD4hiCD8hi gate was used for Cre+ and Cre- sorting, to assure analysis of population expressing similar CD4 and CD8 levels. Independent sorts from two mice per genotype were done: Cre+ A, Cre+ B, Cre- A, Cre- B. Total RNA was extracted for array analysis. Two comparisons were performed: Cre+ to a mouse reference sample and Cre- to this same reference, each done in duplicate (2 biological replicas).