Extended Data Fig. 5: EZH2 CXC-mutants elevate PRC2 methyltransferase activity in vitro and in cells.
From: Drug addiction unveils a repressive methylation ceiling in EZH2-mutant lymphoma
a, Immunoblot showing EZH2 levels in K562 wild-type, CXCdel+/+/+ and Q575R. b, Line plot depicting cumulative growth of K562 wild-type and CXCdel+/+/+ over 10 days. c, Immunoblot showing H3K27me1/2/3 levels in K562 wild-type and CXCdel+/+/+ treated with 5 μM GSK343 or EED226 for 72 h. d, Schematic showing genotypes and allele frequencies for mutations that are observed at frequencies of > 2% in the gDNA encoding the EZH2 CXC ___domain in K562 CXCdel+/+/+. e, Same as d. but for K562 Q575R. f, Same as d. but for Karpas-422 Q575R. g, Purification of recombinant 5-member PRC2 complex. SDS-PAGE stained with Coomassie blue showing purified wild-type, CXCdel, and Q575R PRC2 complexes. h, Representative EMSA showing binding of wild-type, CXCdel and Q575R PRC2 to 185-bp mononucleosomes. The upper band corresponds to the PRC2-bound mononucleosome and bottom band corresponds to the free, unbound mononucleosome. Results in a,c-g are representative of two independent experiments, h is representative of five independent experiments.
