MOESM1 of The cohesin-associated protein Wapal is required for proper Polycomb-mediated gene silencing

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Additional file 1: Figure S1. A) Cells expressing EGFP under the control of the Oct4 locus were generated (see Materials and Methods section for details). GFP mean fluorescence intensity (MFI) was measured 2, 4, and 6 days post-infection. All data are normalized to the GFP MFI of cells infected with the empty vector on day 0, displayed on the y-axis. B) Wild-type (WT) or cells expressing a full-length Wapal cDNA immune to shRNA #2 were infected and bright field (10x, top panel) or alkaline phosphatase stained (4x, bottom panel) are shown 6 days post-infection. Domain structure of Wapal is shown to right of images. C) mRNA levels of different cohesin (Wapal and Smc3) and pluripotency (Nanog, Oct4) are shown. A linear scale of relative expression (compared to WT cells infected with the empty vector) is shown on the y-axis. * indicates a statistically significant reduction from the WT+empty vector control (p value<0.05). D) mRNA levels of differentiation markers. A Log10 scale (left) or linear (right) of relative expression (compared to WT cells infected with the empty vector) is shown on the y-axis. * indicates a statistically significant increase from the WT+empty vector control (p value<0.05). + indicates a statistically significant reduction in expression from WT+Wapal shRNA #2 (p-value<0.05). E) Western blots on cells six days after infection. Antibodies used are indicated on the right. E=empty vector, W= Wapal shRNA #2, WT= wild-type.

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