Fig. 10: GSK-3 Inhibition and Loss of Xist RNA Coating in Female mEpiSCs.
From: Preventing erosion of X-chromosome inactivation in human embryonic stem cells
a Strategy to test the impact of GSK-3 inhibition on Xist RNA coating in mEpiSCs. b Representative RNA FISH images of colonies with high, intermediate, and low percentage of Xist RNA coating (green). Nuclei are stained blue with DAPI. At least ten nuclei were counted per colony in mEpiSC RNA FISH quantification. Scale bars are ~100 microns. c Percentage of nuclei with Xist RNA coating in three independent mEpiSC lines cultured with and without the GSK-3 inhibitor CHIR99021 (CHIR; 3 μM). All three mEpiSC lines cultured with CHIR lost a significant proportion of Xist RNA coating compared to mEpiSCs cultured without CHIR (general linear model comparison, p < 0.001). d Model of direct or indirect repression of human XIST and mouse Xist expression. Conserved mouse and human Xist/XIST sequences upstream of the Xist/XIST TSSs shown, with putative TCF binding motifs in red and surrounding conserved sequence in blue. At least ten nuclei were counted per colony for mEpiSC RNA FISH quantification. The total number of colonies quantified at each passage range between 15 and 25 and are cataloged in source data. Source data are provided as a Source Data file.
