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Fig. 3
Combination of Roblitinib and CPI-169 synergistically inhibits the HCC cell growth. A-C Cell viability of HepG2 (A), SMMC-7721 (B) and MHCC97H (C) cell lines was evaluated by the CCK-8 following increasing concentrations of CPI-169, Roblitinib or CPI-169 + Roblitinib treatment for 48 h. Data are presented as mean ± SEM (n = 3). D Drug interaction analysis between CPI-169 and Roblitinib in HepG2, SMMC-7721, MHCC97H, MHCC97L and Huh7 cell lines. The CI values less than 1.0, approximately 1.0, and greater than 1.0 indicate synergism, additive, and antagonism, respectively. E EdU assay of HepG2 cells following CPI-169, Roblitinib or CPI-169 + Roblitinib treatment for 48 h. Scale bars: 50 ?m. F Colony formation assay of HepG2 and SMMC-7721 cell lines following CPI-169, Roblitinib or CPI-169 + Roblitinib treatment for 14 days. Scale bars: 1 cm. G Measurement of the cell numbers in (E). Data are presented as mean ± SEM (n = 3, one-way ANOVA with Tukey?s multiple comparison test, *p < 0.05, ***p < 0.001, ****p < 0.0001, ns, no significance). H Measurement of the clone numbers in (F). Data are presented as mean ± SEM (n = 3, two-way ANOVA with Sidak?s multiple comparison test, ****p < 0.0001, ns, no significance). I Western blot analysis of the indicated protein expression in HepG2 cells following CPI-169, Roblitinib or CPI-169 + Roblitinib treatment for 48 h. J-K Zebrafish harboring KRASG12V+ HCC primary tumors (J) and zebrafish HCC xenografts using the mCherry-labbled HepG2 cells (K) treated with CPI-169, Roblitinib or CPI-169 + Roblitinib for 72 h. Scale bars: 100 ?m. Data are presented as mean ± SEM (n = 15, one-way ANOVA with Tukey?s multiple comparison test, *p < 0.05, ***p < 0.001, ****p < 0.0001, ns, no significance)