(a) Hypothesis: Tethered G-tract RNA can reverse the ectopic recruitment of PRC2 to specific genes that occurs during cell transformation.
(b) Left: SUZ12 binding to Smad6 nascent pre-mRNA before and after HRasV12 expression, measured by RIP-qPCR with and without UV-crosslinking of cells (mean and S.D., n=3 independent RIPs. Smad6 SUZ12 +/-Ras P=0.043, Welch’s one-tailed t-test). Right: SUZ12, H3K27me3 and total H3 chromatin occupancy (with IgG control) at Smad6 before and after HRasV12 expression (mean and S.D., n=3 independent ChIPs. SUZ12 P=0.036, H3K27me3 P=0.021, Welch’s one-tailed t-test).
(c) Change in HA-dCas9, SUZ12, H3K27me3 and total H3 occupancy at Smad6 and Pax7 after dox-mediated induction of HA-dCas9 in HRasV12-expressing cells containing sgRNA specific for Smad6 (mean and S.D., n=3 independent dox inductions. G-tract RNA: SUZ12 P=0.0082, H3K27me3 P=0.02, Welch’s one-tailed t-test).