Fig. 8

ADAM9 silencing impairs proliferation and invasiveness and increases drug sensitivity of dabrafenib-resistant cells. a Melanoma cell lysates were analyzed by immunoblotting using antibodies against ADAM9 and PIK3R2, or against β-tubulin as a loading control. The results are representative of three independent experiments. b Melanoma cells were transfected with 50 nM pre-miR-126-3p (pre-126-3p) or pre-miR-CTRL and assayed for ADAM9 and PIK3R2 expression 72 h after transfection. c Melanoma cells were transfected with 10 nM (SK-Mel28R) or 50 nM (A375R) siADAM9 or siCTRL and assayed for ADAM9 and PIK3R2 expression 72 after transfection. d, e Melanoma cells were transfected as in (c) and 24 h later incubated with graded concentrations of dabrafenib (DAB) or with DMSO alone. After five days of culture, proliferation was assessed by the MTT assay. Data are expressed in terms of absorbance at 595 nM. Each value represents the arithmetic mean of five independent experiments performed with triplicate samples. Bars, SEM. ^##P<0.01 and ^#P<0.05, siADAM9 vs matched siCTRL; ^**P<0.01 and ^*P<0.05, siADAM9/DAB vs siADAM9/DMSO; ^§§P<0.01 and ^§P<0.05, siCTRL/DAB vs siCTRL/DMSO. f, g Melanoma cells were transfected as in (c) and 72 h after transfection tested for ECM invasion. Left panels, number of invaded cells per microscopic field. Each value represents the arithmetic mean of four (A375R) or five (SK-Mel28R) independent experiments performed with triplicate samples. Bars, SEM. ^**P<0.01 and ^*P<0.05, siADAM9 vs siCTRL. Right panels, representative images of polycarbonate filters with invaded melanoma cells.