Regulation of histone H3 by the ubiquitin-conjugating enzyme UBE2K determines neurogenesis of human embryonic stem cells [ChIP-Seq]

We find that loss of UBE2K leads to accumulation of histone leading to increase in overall load for epigenetic marks and thus hindering differentiation.Our results provide a novel link between ubiquitination, histone regulation and hESC differentiation into neurons. To test whether UBE2K knockdown also induces enrichment of H3K9me3 marks at specific genes, we performed chromatin immunoprecipitation (ChIP)-sequencing assays of hESCs using an antibody to H3K9me3. Overall design: We performed chromatin immunoprecipitation (ChIP)-sequencing using H3K9me3 antibody (Abcam, #8898, reported suitable for ChIP) comparing H9 hESCs expressing non-targeting (NT) shRNA with H9 hESCs expressing UBE2K shRNA.