Opposing functions of H2BK120 ubiquitylation and H3K79 methylation in the regulation of pluripotency

Phf5a regulates transcription elongation in mouse embryonic stem cells (ESCs), through regulation of the Paf1 complex. In this study we assayed for genome-wide localization of H2BK120-ub in mouse ESCs under conditions of shControl and shPhf5a knockdown. These results revealed that downregualtion of Phf5a results in the increase of the elongation mark H2BK120-ub from the gene bodies of pluripotency genes in ESC. Overall design: Ten million cells were treated with micrococcal nuclease to generate mono-nucleosomes and chromatin was precipitated using 10 micrograms of antibody against H2Bk120-ub, clone 56 (Millipore, 05-1312).