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. 2015 Jul 31;6(29):28296–28311. doi: 10.18632/oncotarget.5064

Figure 1. Effects of fisetin on cell viability, colony formation, apoptosis and on modulation of PI3K signaling pathway in BRAF-mutated melanoma cells.

Figure 1

BRAF-mutated melanoma cells (A375, SK-MEL-28 and RPMI-7951) were treated with the indicated concentrations of fisetin. A. The MTT assay was performed to determine the cell viability after 48 hrs of treatment. Data shown here are mean ± SEM of three separate experiments in which each treatment was repeated in 10 wells. *P < 0.05; **P < 0.01 versus control. B. & C. After treatment with fisetin for 24 hrs, the colony assay was performed by seeding melanoma cells in 6-well culture plates at a density of approximately 500 cells/well in 3 ml medium. Cells were allowed to grow in complete growth medium for 2 weeks before crystal violet staining. The data shown here are from a representative experiment repeated three times with similar results. *P < 0.05; **P < 0.01; ***P < 0.001 versus control. For Western blotting, after treatment with fisetin for 48 hrs, the effects on D. cleaved caspase-3, PARP and Bcl2 family proteins, E. modulation of PI3K signaling pathway was determined. Equal protein loading was confirmed by stripping the immunoblot and reprobing it for β-actin. The data shown here are from a representative experiment repeated three times with similar results.