Effect of c-Myb Deficiency in Irf4/8-/- large pre-B cells

Differential gene expression analysis comparing c-Myb-sufficient and -deficient Irf4/8-/- large pre-B cells. The results of this study are the first to demonstrate a role for c-Myb during the large pre-B cell stage of B lymphocyte development. The results provide insight into roles for c-Myb in regulating components of glucose metabolism in large pre-B cells as well as the differentiation of large pre-B cells to the small pre-B cell compartment. Overall design: Irf4/8-/- large pre-B cells were transduced with retroviruses that co-produce a GFP reporter and a Myb mRNA targeting shRNA (shMyb) or a a control Luciferase mRNA targeting shRNA (shLuc) in duplicate. Transduced cells were electronically sorted based on GFP expression and total cellular RNA was prepared. Next generation sequencing (RNA-seq) was performed on isolated mRNA from each sample on the Illumina NextSeq 500 for paired-end sequencing.