Figure 5. Compound 13i HCl increases MLKL phosphorylation and ROS production in bladder cancer cells. (A, B) RT112 cells were treated with the indicated concentrations of 13i HCl (A) or PF-670462 (B) for 24 h, 48 h and 72 h. The cells were subjected to Western blotting with the indicated antibodies. (C) RT112 cells were exposed to 13i HCl for 72 h and LDH levels in the supernatants were determined. (D, E) RT112 cells were exposed to the indicated concentrations of 13i HCl for 72 h or H2O2 (10 μM) for 1 h. The cells were stained with H2DCFDA (D) or MitoSOX (E) and subjected to flow cytometry. Data are represented as mean ± S.D. *P<0.05, **P<0.01, ***P<0.001 compared to control cells (n=3).