Fig. 4

ASC-J9® (ASC) alters cell cycle profiles and apoptotic cell population of BCa cells treated with Cisplatin (Cis). a-b Cell cycle profile and apoptotic cell population in J82 (a) and TCC-SUP (b) cells treated with Cis with/without 5 μM ASC. Cell cycle and apoptosis analyses were assessed by 2 μg/ml DAPI staining for 10 min at room temperature followed by flow cytometry. Apoptotic cells were determined by evaluating the percentage of events accumulated in the subG1 position. c-d The Apoptosis TUNEL Assay was used to determine the apoptosis affected by Cis, ASC, or combined treatment in J82 (c) and TCC-SUP (d) cells. e-f Apoptosis signals of BCL-2 and BAX affected by treatment with Cis, ASC or both were evaluated by RT-PCR (e) and by Western blotting (f) assay. g-h The cell proliferation BrdU incorporation assay and cell proliferation regulatory proteins were affected by Cis, ASC or combined treatment in J82 (g) and TCC-SUP (h) cells. i Cell growth inhibitory signal p21 RNA affected by treatment with Cis, ASC or both on J82 and TCC-SUP cells were evaluated by RT-PCR. j Cell growth inhibitory signal of p21 proteins affected by treatment with Cis, ASC or both on J82 (upper) and TCC-SUP (lower) cells were evaluated by Western blotting assay. For (f) and (j) the relative p21 expressions (the number under the bands) were determined by measuring densities of corresponding bands on western blots and normalizing to β-ACTIN. Data are presented as mean ± SD. *, p < 0.05; **, p < 0.01