Additional file 6: of Kinase inhibitor library screening identifies synergistic drug combinations effective in sensitive and resistant melanoma cells

Figure S4. Western blot analysis for selected drug treatments and apoptosis assays in healthy and melanoma cells. A) Western Blot analysis of A375, A375-XP and A375-GP cells treated with the BRAFi Vemurafenib (PLX), Chki AZD7762 (AZD), Wee1i MK-1775 (MK), FAKi TAE226 (TAE) or combinations thereof. Cells were treated for 3 h with indicated concentrations of inhibitors. Actin staining was used as loading control. B) The combination of MK-1775 and AZD7762 efficiently induced apoptosis in primary melanoma cells (M45), but not so much in healthy cells. Cells were treated for 72 h with the indicated concentrations of MK-1775 (Wee1i) or AZD7762 (Chki) or a combination thereof. Etoposide (Eto) treatment was used as positive apoptosis control. Resulting caspase-3 activity was normalized to the untreated control. 1 representative experiment out of 3 is shown. C) Western blot analysis of NHEM, NHDF and M45 primary melanoma cells after treatment for 3 or 24 h with indicated amounts of drugs. P-cdc2 (CDK1), cdc2 (CDK1), p-Chk1 and Chk1 were detected after 3 h drug treatment, while PARP cleavage was detected after 24 h treatment. Vinculin and α-tubulin were used as loading controls. AZD: AZD7762, MK: MK-1775; NHEM. Normal human epidermal melanocytes, NHDF: normal human dermal fibroblasts. (PDF 306 kb)