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Fig. 3 | Acta Neuropathologica Communications

Fig. 3

From: DNA hypomethylator phenotype reprograms glutamatergic network in receptor tyrosine kinase gene-mutated glioblastoma

Fig. 3

mTORC2 redistributes EZH2 in the DNMT3A promoter to suppress its expression. A Immunoblot detection of DNMT3A and H3 p.K27me3 in U87-EGFRvIII cells treated with GSK126 (EZH2 inhibitor: 2.5 µM) and GSKJ4 (JmjC inhibitor: 10 µM) for 48 h. B, C ChIP-qPCR analysis on H3 p.K27me3 (B) and EZH2 (C) enrichment in DNMT3A promoter regions of U87-EGFRvIII cells transfected with siRNAs against Scramble sequence or Rictor. D Immunoblot analyses of acetylated EZH2 (Ac-EZH2) in U87-EGFRvIII cells with shScramble or shRictor. Ac-K, acetylated-lysine; IB, immunoblotting; IP, immunoprecipitation. E Immunoblot analyses of acetylated EZH2 (Ac-EZH2) in U87 cells with overexpression of GFP or Rictor. Ac-K, acetylated-lysine; IB, immunoblotting; IP, immunoprecipitation. F Analyses on acetylation and redistribution of EZH2 on the DNMT3A promoter in U87-EGFRvIII cells with addition of TSA (1.0 µM) and acetate (10 mM) for 48 h. Ac, acetate. G mRNA expression of DNMT3A in U87-EGFRvIII cells treated by PP242 (mTORC1/C2 inhibitor: 5 uM) along with supplementation of TSA (1.0 µM) and acetate (10 mM) for 48 h. H mTORC2 drives protein acetylation to redistribute EZH2 into the DNMT3A promoter region, and increases H3 p.K27me3 to suppress the expression of DNMT3A in GBM. Ac, acetyl-group

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