Fig. 9: GSK-3 Inhibition and Loss of Xist RNA Coating in Differentiating Female mESCs.
From: Preventing erosion of X-chromosome inactivation in human embryonic stem cells
a Strategy for the differentiation of mESC lines into mEpiLCs and culture of mEpiLCs with and without the GSK-3 inhibitor CHIR99021 (CHIR; 3 μM). b Representative images of mEpiLCs with high, intermediate, and low percent of nuclei with Xist RNA coating (green). Nuclei are stained blue with DAPI. At least ten nuclei were counted per colony in mEpiLC RNA FISH quantification. Scale bars are ~100 microns. c Percentage of nuclei with Xist RNA coating in differentiating mEpiLCs with and without CHIR generated from three independent ESC lines. mEpiLCs cultured with CHIR lost a significant proportion of Xist RNA coating compared to mEpiLCs cultured without CHIR in all three mEpiLC replicates (general linear model comparison, p < 0.001). At least ten nuclei were counted per colony for mEpiLC RNA FISH quantification. The total number of colonies quantified at each passage range between 12 and 20 and are cataloged in source data. Source data are provided as a Source Data file.
