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. 2020 Jul 9;10(18):8098–8110. doi: 10.7150/thno.45363

Figure 3.

Figure 3

Mcl-1 inhibitors re-sensitize CRC cells with acquired resistance to regorafenib. (A) MTS analysis of parental and regorafenib-resistant HCT116 (HCT116-R) and Lim1215 (Lim1215-R) cells treated with regorafenib at indicated concentrations alone or in combination with an indicated Mcl-1 inhibitor (5 μM) for 72 hours. (B) Crystal violet staining of parental and regorafenib-resistant HCT116 and Lim1215 cells treated with regorafenib (40 μM) alone or in combination with S63845 (5 μM) for 48 hours. (C) Colony formation of cells treated as in (B). Upper panel: representative pictures of colonies visualized by crystal violet staining 2 weeks after treatment; lower panel: enumeration of colony numbers. (D) Apoptosis in cells treated with regorafenib (40 μM) alone or in combination with indicated Mcl-1 inhibitor (5 μM) for 48 hours was analyzed by counting condensed and fragment nuclei. (E) Western blotting of Mcl-1 and cleaved (C) caspases 3, 8, and 9 in indicated cells treated as in C. (F) IP analysis of the binding between endogenous Mcl-1 and PUMA in parental HCT116 and HCT116-R cells treated as in C for 24 hours. In A, C and D, results were expressed as means ± s.d. of three independent experiments. NS, not significant, P >0.05; **, P <0.01; ***, P <0.001.