Fig. 3.

BRAFi or MEKi combine with AZD5991 to inhibit melanoma cell survival and tumour growth. a A375 cells seeded at low density were treated with DMSO control (C), 2 μM vemurafenib (Vem), 1 μM AZD5991 or the combination for 72 h. b A375 cells seeded at low density were treated with DMSO control (C), 1 μM selumetinib (Sel), 1 μM AZD5991 or the combination for 72 h. c A375 cells seeded at low density were treated with DMSO control (C), 2 μM vemurafenib (Vem), 1 μM AZD4320 or the combination for 72 h. a–c Following treatment cells were washed and grown in inhibitor free medium for a further four days. Colonies were then visualised by crystal violet staining and counted and then growth assessed following solubilisation. Results are mean ± SD of three or more independent experiments. P ≤ 0.0001 (****), P ≤ 0.05 (*) or ns (not significant) as determined by one-way ANOVA and Sidak’s multiple comparisons test. d, e Female nude athymic mice were implanted subcutaneously with A375 cells and randomised 21 days later for dosing with either vehicle-only (Control, n = 5), 25 mg kg⁻¹ selumetinib (Sel, n = 7) twice daily with an 8 h interval, 60 mg kg⁻¹ AZD5991 (n = 7) intravenously once weekly, or the combination of 25 mg kg⁻¹ selumetinib and 60 mg kg⁻¹ AZD5991 (Sel + AZD5991, n = 8) for a further 21 days. Tumour growth was recorded twice weekly and results are mean ± SEM (d) or mean % change in tumour volume ± SEM (e). P ≤ 0.0001 (****), P ≤ 0.001 (***) or P ≤ 0.05 (*), as determined by two-way ANOVA and Holm-Sidak’s multiple comparisons test