Fig. 5. THZ1 represses MCL1 expression and synergizes with ABT-263 in CCA cell lines.

a Time-dependent effect of THZ1 200 nM and dose-dependent effect of THZ1 12 h on mRNA level of MCL1 were measured by RT-qPCR in indicated cells. Data represent mean ± SEM of three independent replicates. b Time-dependent effect of THZ1 200 nM and dose-dependent effect of THZ1 24 h on protein levels of MCL1, BCL2, and BCL-XL were analyzed by western blotting. c The expression level of CDK7 showed a significant positive correlation with MCL1 expression in the tissue microarray. The r and P values were determined by Spearman correlation analysis. d The indicated cells were treated with combination of three concentrations of THZ1 and ABT-263 for 48 h. The heatmaps show the results of percent cell viability of three independent results. CI was determined using CompuSyn software. e HuCCT1 and HuH28 cells were treated with THZ1 and ABT-263 at indicated concentrations for 48 h. The apoptosis was detected by the Annexin-V/PI assay and Caspase 3/7 activity. Data represent mean ± SEM of three independent replicates (**P < 0.01; ***P < 0.001). f HuCCT1 and HuH28 cells were treated with THZ1 and ABT-263 at the above-mentioned concentrations for 48 h. Cleaved PARP protein expression was analyzed by western blotting