Research Paper Volume 12, Issue 7 pp 5764—5780

Figure 4. Inhibition of CK1δ activates necroptosis in bladder cancer cells. (A) RT112 cells were treated with the indicated concentrations of 13i HCl in the presence or absence of zVAD (20 μM) or NSA (10 μM) for 48 h and subjected to MTT assay. Data are represented as mean ± S.D. **P<0.01 compared to the group of 13i HCl alone (n=3). (B) RT112 cells were treated with the indicated concentrations of 13i HCl in the presence or absence of zVAD (20 μM) plus NSA (10 μM) for 48 h and subjected to MTT assay. Data are represented as mean ± S.D. *P<0.05, **P<0.01 compared to the group of 13i HCl alone (n=3). (C, D) RT112 and MLKL stable knocked-down clone (shMLKL-1, shMLKL-2) cells were treated with the indicated concentrations of 13i HCl (C) or PF-670462 (D) for 72 h and subjected to MTT assay. Data are represented as mean ± S.D. *P<0.05, **P<0.01, ***P<0.001compared to the group of 13i HCl or PF-670462 alone (n=3). (E) The knockdown efficiency of MLKL in RT112 cells was examined by Western blotting.