Fig. 5

In vitro cytotoxic effects of ITC-6102RO through apoptosis. (A) H23 and JIMT-1 cells were treated with Naked mAb, ITC-02-050 (free drug), or ITC-6102RO at various concentrations (0, 0.001 ~ 0.1 µM) for 48 h. Each point represents the mean and SD (n = 8). After incubation, cellular viability was measured using the cell counting kit-8. Data are shown as mean ± standard deviation. ***p < 0.001 versus Naked mAb-treated cells. (B) Proteolytic processing of caspase-3 by ITC-6102RO. H23 and JIMT-1 cells were treated with Naked mAb, ITC-02-050 (free drug), or ITC-6102RO at 0.1-µM for the indicated times. Cell extracts were prepared for Western blotting to detect changes in the expression of caspase-3 and PARP-1. Equal loading of protein samples was verified using Western blotting of α-tubulin. (C) The apoptotic effects of ITC-6102RO in H23 and JIMT-1 cell lines treated with Naked mAb, ITC-02-050 (free drug), or ITC-6102RO at 0.1-µM for 48 h. Each cell line was stained using Annexin V-PI and examined by flow cytometry as described in the Materials and Methods. The results presented are mean ± SD, with n = 3 replicates in each group. **p < 0.01, ***p < 0.001 versus Naked mAb-treated cells