H2AK119ub1 native ChIP-seq upon Xist indution

Native ChIP-seq for H2AK119ub1 upon Xist induction in iXist-ChrX mouse embryonic stem cells. Comparison of WT with Spen RRM deletion and SPOC domain mutants for two timepoints of Xist induction, 3h and 24h. ChIP-seq for H2AK119ub1 was performed on native chromatin extract from WT and biological replicate clones for Spen RRM deletion and SPOC mutation cell lines derived from parental iXist-ChrX cells. For the purposes of this report H2AK119ub1 distribution acts a surrogate marker for Xist RNA distribution across the Xi chromosome, and its pattern is clearly abnormal in Spen RRM deletion but not SPOC mutant cells. Two time points of Xist induction, 3h and 24h were profiled for comparison. Whereas all samples for the 3h experiment were processed in parallel on the same day, for the 24h experiment the Spen RRM deletion lines, SPOC mutant lines, and each of the three WT replicates were processed on separate occasions. For 24h data sets (WT Reps1&2, SpenRRM, SPOCmut), Drosophila SG4 cells were carefully added as a spike in (40 million mES cells with 10 million SG4 cells) upon collection and prior to chromatin extraction. Although spike-in normalisation was not performed for this study, input samples are provided here for this analysis.