The role of AF10 (Mllt10) in maintaining cell identity (ChIP-Seq)

AF10 is a cofactor of the H3K79 methyltransferase DOT1L. To uncover the role of H3K79me in reprogramming to induced pluripotent stem cells (iPSCs), we bred reprogrammable mice encoding doxycycline inducible Oct4-2A-Klf4-2A-IRES-Sox2-2A-c-Myc (OKSM) transgene with conditional flox (fl)-AF10 (Mllt10). We isolated mouse embryonic fibroblasts (MEFs), deleted AF10 with CRE-recombinase or treated cells with empty vector control, and initiated reprogramming in DOT1L chemical inhibitor SGC0946 or DMSO control. Gene expression was assessed using RNA-Seq and genome wide localization of H3K79me1, H3K79me2, and RNA Polymerase II (RNAPII) was determined by ChIP-Seq on day 4 of reprogramming. Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for H3K79me1, H3K79me2, and RNA Polymerase II (RNAPII) in cells on day 4 of reprogramming to iPSCs with wild-type (fl) or Cre-deleted AF10 (Mllt10)