Fig. 5

Effect of combinatorial treatment of parental FGFR3 fusion harboring bladder cell lines with an FGFR and pan-ERBB inhibitor on (A, E) cell signaling, (B, F) cell viability, (C, G) cell cycle kinetics and (D, H) apoptosis. A, E Parental SW780 and RT4 cells cells were treated with BGJ398 (0.1 µM) or AZD8931 (1 µM), alone or in combination for 72 h and changes in pERBB3 and pERK determined by western blot. B, F Parental SW780 and RT4 cells were treated with a range of concentrations of BGJ398 or AZD8931 alone or in combination for 72 h and cell viability determined using Cell-Titer Glo assays. Plots shown are the BLISS synergy analysis from a representative experiment, which shows synergistic growth inhibition across a range of concentrations. C, G FGFR inhibitor-resistant (C) SW780-RS and (G) RT4-RS cell lines were treated with BGJ398 alone and in combination with the pan-ERBB inhibitor, AZD8931, for 24 h and changes in cell cycle distribution determined by propidium iodide staining and FACS analysis. D, H Assessment of the effect of combination treatment with BGJ398 and AZD8931 on apoptosis by propidium iodide staining and FACS analysis in the same samples analysed in panels C and G. Values shown are mean ± SEM of a representative experiment performed in triplicate. *P < 0.05 and ***P < 0.0005, t test