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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Cancer Res. 2018 May 29;78(15):4360–4369. doi: 10.1158/0008-5472.CAN-17-3124

Figure 2. The combined treatment reduces cell viability through multiple mechanisms.

Figure 2

(A–B) The combination of CDK4/6 and c-Met/Trk inhibition suppresses the cell cycle. Protein expressions were detected by immunoblot using specific antibodies for p-Rb and Cyclin A, quantitated and plotted (*P < 0.05; ***P < 0.0001; one-way ANOVA with post-hoc Tukey analysis). Shown also is cell cycle analysis of G88 cells following 24 hours of treatment with abemaciclib, altiratinib, and the combination of both (*P < 0.05; **P < 0.01 (control vs. treatment groups); two-tailed t-test). (C–D) The combined treatment induces significant apoptosis. Shown is an immunoblot using antibody specific for PARP. Protein expressions were quantitated and plotted (***P < 0.0001; one-way ANOVA with post-hoc Tukey analysis). In addition, caspase-3/7 activity is significantly increased with two days of combined treatment. (***P < 0.0001; one-way ANOVA with post-hoc Tukey analysis). (E) The pan-caspase inhibitor, Z-VAD-FMK (25 μM), partially reversed cytotoxicity from the combined treatment. Cell viability was assessed at 72 hours of treatment (***P < 0.0001, one-way ANOVA with post-hoc Tukey analysis). NS: Non-significant. All values are mean ± SEM and from three biologically independent samples. The concentrations of abemaciclib and altiratinib used in the experiments were 1.5 μM and 2.5 μM, respectively.