Fig. 6. Contribution of PRC2 dimerization to H3K27me3 on chromatin in vivo.

(A) Western blot confirming SUZ12 expression levels in SUZ12^WT and SUZ12^3D mESCs grown in the 2i condition, using an anti-SUZ12 antibody. Global H3K27me3 levels were detected. Anti-GAPDH signals were used as loading controls.

(B) SEC profiles for endogenous EZH2 (upper panel) and MTF2 (lower panel) in SUZ12^WT and SUZ12^3D mESCs. Elution profile was depicted according to Western blot signals. Positions of a 670KD (Bio-Rad SEC standard) and a 200KD (molecular weight of the SUZ12(N)-RBBP4 dimer) marker protein are indicated by arrows.

(C) The same as (A), for SUZ12^WT and SUZ12^3D HEK293T cells.

(D) H3K27me3 enrichment by ChIP-qPCR on gene loci not targeted by PRC2. In (D), (E), (F) and (G), SUZ12^WT and SUZ12^3D mESCs grown in three different days were used. Graphs display mean ± SEM from a total of six ChIP-qPCR reactions. GraphPad Prism 8.0 was used for data analysis. P values were derived from two-tailed Student’s t-test: ns (p>0.05); * (p≤0.05); ** (p≤0.01); *** (p≤0.001); **** (p≤0.0001).

(E) H3K27me3 enrichment on PRC2 targets in mESCs grown under the 2i condition.

(F) and (G) The same as (D) and (E), except that mESCs were grown under the serum condition.

See also Fig. S6 and Table S1.