Figure 6 | Scientific Reports

Figure 6

From: Fisetin targets YB-1/RSK axis independent of its effect on ERK signaling: insights from in vitro and in vivo melanoma models

Figure 6

Comparable inhibition of melanoma tumor growth by fisetin and vemurafenib albeit through divergent regulation of ERK signaling (A) Whole cell lysates of A375 melanoma cells treated with fisetin (60 µM) or vemurafenib (10 µM) for 24 h were analyzed for phosphorylated RSK, ERK1/2, total YB-1 and MDR1. Equal loading was confirmed by reprobing for vinculin. (B) Kinase inhibitory activity for ERK1/2. Fisetin was tested in 10-dose IC50 mode with 3-fold serial dilution starting at 100 μM. Reactions were carried out at 30 μM ATP. (C) Whole cell lysates of A375 melanoma cells treated with fisetin (60 µM) or vemurafenib (10 µM) for specified time points analyzed for phosphorylated and total ERK1/2. Equal loading was confirmed by reprobing for vinculin. (D) Whole cell lysates of A375 melanoma and NCI/ADR-Res ovarian cancer cells, treated with fisetin (60 µM) and/or ERK1/2 inhibitor FR180204 (10 µM) for 24 h were analyzed for cleaved caspase-3 expression. Equal loading was confirmed by reprobing for vinculin. (E) Average tumor volume of DMSO/fisetin/vemurafenib-treated mice plotted over days after tumor cell inoculation. Points, mean of 12 tumors in six animals; bars, mean ± SE,*p ≤ 0.05, **p ≤ 0.01 versus the control group. (F) Whole cell lysates of tumor tissues analyzed by western blot. Equal loading was confirmed by reprobing for GAPDH. Data represent samples from each group repeated twice with similar results.

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