Figure 2.

The effects of co-treatment with thioridazine plus TRAIL on apoptosis in other carcinoma cell lines, and normal cells. (a) Renal carcinoma (ACHN and A498), breast carcinoma (MDA-MB231), and glioma (U251MG) cells were treated with 50 ng/ml TRAIL in the presence or absence of 10 μM thioridazine for 24 h. The level of apoptosis was measured by the sub-G1 fraction using flow cytometry. The protein expression levels of PARP and actin were determined by western blotting. The level of actin was used as a loading control. (b) Caki, TMCK-1, and mesangial cells were treated with 50 ng/ml TRAIL in the presence or absence of 10 μM thioridazine for 24 h. The cell morphology was examined using interference light microscopy (left panel). The level of apoptosis was analyzed by measuring the sub-G1 fraction by flow cytometry (right panel). (c) Renal carcinoma (ACHN and A498), breast carcinoma (MDA-MB231), and glioma (U251MG) cells were treated with the indicated concentrations of thioridazine for 24 h. The protein expression levels of c-FLIP(L), Mcl-1, and actin were determined by western blotting. The level of actin was used as a loading control. The values in (a and b) represent the mean±S.D. from three independent samples. *P<0.001 compared with the thioridazine treatment alone. The data represent three independent experiments