Fig. 6

DNA demethylation induces Xi reactivation upon reprograming at loci within high H3K9me3 and low H3K27me3 domains. a Scheme illustrates the experimental outline. hF clones were cultured in the presence of 5-azaC for three days before fusion with mESCs. 5-azaC was removed from cultures at fusion and allelic expression of X-linked genes was analysed 0, 4 and 6 days post fusion. b Histogram plots show Xi-allelic expression during reprograming upon 5azaC pre-treatment. Inactive alleles with a significant Xi induction upon 5azaC pre-treatment and cell fusion-mediated reprograming are shown in orange, whereas refractory alleles are shown in red. Asterisks (*) mark significant changes vs. day 0 (p ≤0.05, two-sided t-test). Allelic expression was measured by Taqman probes as NRFU of the Xi-specific probe versus the total. Data represent the average of two independent experiments ± SEM for each clone. c Box plots show Xi-specific enrichment of H3K9me3 and H3K27me3 in clones 12 and 34. Xi enrichment has been detected by using SNP-specific Taqman probes and normalising Ct values to H3 ChIP and input DNA. ChIP was performed in at least two independent experiments for each clone