Fig. 4: SPEN plays a role in Xist RNA stability.
From: SPEN is required for Xist upregulation during initiation of X chromosome inactivation
a Relative total Xist expression in undifferentiated Wt and Spen−/− ESCs upon Xist induction with doxycycline for 4 days. n = 3 biological replicates per condition. Average expression ± SD. Reference gene: Beta-actin. b Xist RNA FISH (red) of undifferentiated Wt and Spen−/− ESCs treated with doxycycline to induce Xist expression. DNA is stained with DAPI (blue). Scale bar: 10 µm. c Quantification of (b), displaying the percentage of nuclei with Xist clouds. Average percentage, n = 2 biological replicates per condition, the total number of counted nuclei is indicated. d Percentage of Xist RNA remaining at different time points of actinomycin D treatment (t = 0, 2, 6 and 10 h) in undifferentiated Wt and Spen−/− ESCs treated with doxycycline. Normalized relative expression to t = 0 ± SD, shading represents the SD, n = 3 biological replicates per condition. Reference gene: Beta-actin.
