GSH de novo synthesis support cellular physiology in IDH1-mutated cells
A. Western blot measures the expression of GSH synthesis enzymes in U251 cells with IDH1 mutant expression. β-actin was used as internal control.
B. Quantitative real-time PCR analysis measures mRNA level of GSH synthesis enzymes. *p<0.05, **p<0.01.
C. GSH and GSSG level was measured in IDH1-mutated U251 cells. Catalase (Cata) was used as exogenous ROS scavenger (500U/mL, 24 hr). **p<0.01.
D. Annexin V/PI apoptotic analysis in IDH1-mutated U251 cells with genetic silencing of GCLC and GCLM.
E. Quantification of apoptotic cells in D.
F. ROS-Glo assay in IDH1-mutated U251 cells with genetic silencing of GCLC and GCLM.