Fisetin inhibits YB-1 in BRAF mutant melanoma cells (A) Whole cell lysates of A375 melanoma cells treated with fisetin (20–60 µM:24 h) were analyzed for p-YB-1 and YB-1 protein expression. Equal loading was confirmed by reprobing for vinculin. (B) Representative micrographs (20x) showing immunofluorescence for YB-1 in 451Lu melanoma cells with/without fisetin (60 µM:24 h). Scale bar, 20 µm. (C) WM35 melanoma cells transfected with pcDNA-HA-YB-1 and treated with fisetin (60 µM:24 h) were analyzed for YB-1 expression. (D) Histogram represents relative YB-1 mRNA levels in fisetin-treated A375 cells (20–60 µM:24 h) normalized to GAPDH. (E) Representative micrographs (10x) showing YB-1 expression in A375 melanoma constructs treated with fisetin (80 µM), harvested at day 12 and 16 post treatment. Scale bar, 20 µm.