Extended Data Figure 5: SHARP, LBR, SAF-A, SMRT, and HDAC3 are required for Xist-mediated transcriptional silencing.
From: The Xist lncRNA interacts directly with SHARP to silence transcription through HDAC3
a, Representative images showing staining of DAPI (blue), Xist (red), and Gpc4 (green) for different siRNA knockdown in male ES cells before Xist induction (−Dox; left) or after Xist induction for 16 h (+Dox; middle and right). b, Quantification of the copy number of Gpc4 in –Dox and +Dox cells after knockdown with siRNAs targeting different mRNAs. Error bars represent the standard error of the mean across 50 individual cells from one experiment. NS, not significantly different between +Dox and –Dox cells; ****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. c, Knockdown of SHARP, LBR, or SAF-A abrogates Xist-mediated gene silencing without causing pluripotency defects. Representative images showing staining of Nanog (cyan), Xist (red), and Gpc4 (green) upon knockdown of SHARP, LBR or SAF-A after 16 h of Xist induction with doxycycline. Scale bars on the images represent 5 μm.
