Fig 1: Synergistic effects of vemurafenib/PIK-75/miR-126 on the PI3K/AKT and MAPK pathways evaluated in the A375M-TripZ (TripZ) vs A375M/miR-126 (miR) cell lines. Representative western blots illustrate the patterns of AKT and ERK phosphorylation, PARP and cyclin D1 expression in melanoma cells exposed to decreasing concentrations of (A) PIK-75 (PIK; 0.5–0.125 µm) and (B) vemurafenib (Vem; 10–2.5 µm) for 5 h, and the expression levels of (C) AKT and ERK phosphorylation and (D) caspase 3, PARP and cyclin D1 in melanoma cells exposed to vemurafenib (Vem; 5 µm) or PIK-75 (PIK; 0.5 µm) as single agents or in combination for 5 h. (E) Quantification of apoptotic cell fractions in TripZ and miR-126 after 24 h treatment with vemurafenib (Vem; 500 nm) or PIK-75 (PIK; 80 nm) alone or in combination (V + P). Columns, mean ± SD of at least three independent experiments (left panel). Differences were statistically evaluated by Student's t-test, **P < 0.01. Percentages of apoptotic cells in TRIPZ and miR-126 control cells cultured in 10% or 0.1% FBS (right panel). a-Tubulin or ß-actin was used as protein loading control. V + P: vemurafenib + PIK-75.