Fig. 2: Increased levels of Xist RNA silences Xi escapees in astrocytes.
From: Escape from X inactivation is directly modulated by Xist noncoding RNA
a, Experimental outline showing that TX NPCs (E6 clone) carrying a ptet promoter upstream of the Xist gene on the B6 X chromosome were differentiated into astrocytes without Dox. Xist RNA levels were increased by adding Dox to the astrocyte culture media. b, FISH for Xist RNA (green) in the untreated-astrocyte condition (control) and after 3 days of Dox treatment. DNA is stained with DAPI (blue). c, RNA-seq data showing the fold change in Xist expression (normalized CPM) compared with untreated cells across the time course of Dox treatment (17.5–21.1-fold enrichment). Two individual biological replicates are shown per condition. d, Heat map showing the mean allelic ratios between two replicates of escapees after 3 and 7 days of Dox treatment. Escapees are assigned to three different categories as shown in Extended Data Fig. 1g and described in the Methods. The escape category for each gene is indicated below the heat map. e, Scatterplot comparing the allelic ratios of individual escapees in astrocytes and NPCs (E6 clone) for the different Dox treatment conditions. The number of genes matching and Pearson correlation between the two cell types are indicated in the ‘control’ scatterplot.
