IDH2R172 mutant alters histone lysine trimethylation in Jurkat T cells and AITL. (A) Western blots showing ectopic expression of wild-type IDH2 and IDH2R172 in whole cell lysates of transduced Jurkat cells. LC-MS/MS showing alteration of intracellular 2-HG in corresponding groups (n = 4 each, data are shown as mean ± SEM, ***P < .001 by 2-tailed Student t test). (B) Western blots showing changes in trimethylation of various histone lysine residues in transduced Jurkat cells. Histone proteins were extracted and assessed for levels of histone lysine methylation. Total H3 was used as loading control. Blots shown are representative of ≥3 replicates. (C) Representative immunohistochemistry of AITL tissue microarrays with antibody against H3K27me3 (n = 3 in each group). Original magnification, ×40. (D) Image quantification using Image Scope (details in “Materials and methods”). Error bar indicates SEM of 3 independent samples in each group.
