Fig. 2

The regions of XIST critical for histone deacetylation and enrichment of H4K20me1 and CIZ1. A Example images showing typical depletion of H3K27ac (red) at the site of XIST RNA after five days of induction in the HT1080 cell line and z-score s showing the depletion of H3K27ac over time by XIST induction. B Each deletion construct induced for 5 days was tested for its relative depletion (z-score) of H3K27ac at the site of XIST (59–60 cells per construct). The distributions of z-scores for each construct are shown in boxplots with significance calculated relative to the Full XIST control. C Similar to A, example images and time point analysis were performed for the enrichment of H4K20me1 at the site of Full XIST expression in the HT1080 cell line. D Similar to B, the deletion constructs were tested for their enrichment of H4K20me1 after 5 days of XIST induction and the population distributions of the 60 cells analysed are shown in boxplots. E Example images of the enrichment of CIZ1 at the site of XIST and the population distribution of the enrichment profile of cells (z-score) at various induction timepoints. F The plotted enrichment profiles of CIZ1 at XIST in each of the deletion constructs. A–F All samples were blinded throughout the analysis process and statistical significance calculations were performed using the Mann–Whitney test with the thresholds for statistical significance of all p values being: * < 0.05, ** < 0.01 and *** < 0.001, with these thresholds adjusted based on the number of tests