(A) RKO, LOXIMVI, WiDr, A375 and COLO205 cancer cell lines were treated with 1 μM AZD6244, 0.1 μM AZ628, 10 μM PLX4720 or a combination of AZD6244 with either AZ628 or PLX4720 for 18 h. Cell lysates were analyzed by Western blotting for the indicated proteins.
(B) Cell lines were treated as in (A) for 96 h and cell proliferation was assessed by CellTiter-Glo assay.
(C) Cells were treated with a titration of AZD6244 versus a titration of either AZ628 or PLX4720 in combination for 96 h. Cell proliferation was assessed using CellTiter-Glo and presented in the blue-red heat maps. The green-red heat maps present the degree of synergistic interaction between the compounds, as determined using the ‘Bliss Independence model’ where values greater than 0 indicate an effect greater than the combined fractional inhibition of the single agents, indicative of synergy.