Figure 2
Monoallelic upregulation of Xist RNA upon female rESCs differentiation. (a) Xist and Tsix qPCR expression analysis in female (A10p20, A4p20, 135-7, 141-6) and male (A8p20) differentiating rESCs. Expression levels of Xist and Tsix at different time points upon neuronal differentiation are shown. In the Xist panel error bars represent standard deviation of technical replicates from one experiment. In the Tsix panel error bars in A10P20, A4P20 and A8P20 represent standard deviations of technical replicates from one experiment and in 135-7 and 141-6 standard deviations of technical replicates from two experiments. (b) Representative images of Xist RNA FISH (green) analysis upon differentiation of female (A10p20-upper panel) and male (A8p20-lower panel) rESCs. DNA is stained with DAPI (blue). Scale bars represent 10 μm. (c) Quantification of relative number of Xist RNA signals (pinpoints or clouds) in A10p20 (left graph) and A4p20 (right graph) female rESCs at day 0, 2, 3, 4 and 6 upon neuronal differentiation. Xist clouds were never observed in male nuclei. More than 100 nuclei were counted for each time point and for each experiment. Error bars represent standard deviation of two experimental replicates. Asterisks indicate significant difference between the two groups (2 pinpoints versus cloud with or without pinpoint) within each differentiation time point. P-values are provided: p-valueA10-d0 < 0.0001, p-valueA10-d2 = 0.02, p-valueA10-d3 = 0.03, p-valueA10-d4 = 0.03 and p-valueA4-d0 < 0.0001, p-valueA4-d2 = 0.03, p-valueA4-d4 = 0.05, p-valueA4-d6 = 0.03.
