Fig. 2: Spen cDNA expression in Spen−/− ESCs rescues Xist expression.
From: SPEN is required for Xist upregulation during initiation of X chromosome inactivation
a Relative total Xist expression determined by RT-qPCR at days 0, 3, and 5 of monolayer differentiation of Wt, Spen−/− and three independent rescue clones (Clone A, B and C). Average expression ± SD, n = 4 biological replicates per condition. Reference gene: Hist2h2aa1. b RNA FISH of Xist (red) at days 0 and 3 of monolayer differentiation of Wt, Spen−/−, and Spen rescue clones. DNA stained with DAPI (blue). Scale bar: 10 µm. c Quantification of (b), showing the percentage of nuclei with Xist clouds in each condition. Average percentage, n = 2 biological replicates, the total number of counted nuclei is indicated.
