Fig. 4

SCCHN cell lines exposed to a combination of A-1331852 and S63845 (100 nM each) for 24 h exhibited a enhanced apoptosis and b decreased clonogenicity, compared to the individual BH3 mimetics. c Phase contrast images (scale bars 100 μm), and d line graph to show fold change in spheroid volume over 12 days, following exposure to the specified drug(s). Dotted lines in c demarcate the outline of each intact spheroid. e Western blot analysis of antiapoptotic protein levels in SCCHN primary cells. These cells, exposed to a combination of A-1331852 and S63845 (100 nM) for 4 h exhibited f enhanced apoptosis and g decreased clonogenicity, compared to the indicated treatments. a, b, f One-way ANOVAs with Dunnett’s multiple comparisons tests, with a single pooled variance; d two-way repeated measures ANOVA with Dunnett’s multiple comparisons test; g unpaired, two-tailed t tests. Error bars = mean ± SEM of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.