RNA-seq analysis of umbilical cord blood cells upon knockdown of NAP1L3

The nucleosome assembly proteins (NAPs) are histone chaperones with an important role epigenetic regulation of gene expression. We find that high gene expression levels of murine Nap1l3 is restricted to hematopoietic stem cells. Loss of function of murine and human NAP1L3 impair maintenance of hematopoietic stem cells and differentiation both in vitro and in vivo. NAP1L3 inhibition in human hematopoietic cells causes a growth arrest in the G0 phase of cycle progression, and induces gene expression signatures that highly significantly correlate with downregulation of genes involved in cell cycle regulation, including E2F and MYC target genes. In addition, we also show that the HOXA3, HOXA5, HOXA6 and HOXA9 are markedly upregulated when NAP1L3 is suppressed in Human hematopoietic cells. Taken together, our findings establish an important role for NAP1L3 in HSC homeostasis and hematopoietic differentiation. Overall design: Umbilical cord blood cells were transduced with shRNA targeting NAP1L3 or negative control vectors and RNA was isolated for RNA-seq analysis 72hour post transduction.