Extended Data Figure 4: Previously identified proteins associated with XCI are not required for Xist-mediated transcriptional silencing.
From: The Xist lncRNA interacts directly with SHARP to silence transcription through HDAC3
a, To confirm the specificity of our assay, we tested the function of several proteins that were previously identified to associate with Xist, but not to silence transcription, for their role in transcriptional silencing in our inducible male ES cells before Xist induction (−Dox; left) or after Xist induction for 16 h (+Dox; middle and right). Representative images are shown after knockdown of each protein. DAPI (blue), Xist (red), and Gpc4 (green). b, Quantification of the copy number of Gpc4 before and after Xist induction upon treatment with different siRNAs. Error bars represent the standard error of the mean across 50 individual cells from one experiment. ****P value < 0.001 between +Dox and –Dox cells based on an unpaired two-sample t-test. Scale bars on the images represent 5 μm. Importantly, while these proteins do not have a role in the initiation of transcriptional silencing, we do not mean to imply that they do not have other roles in XCI.
