Fig 2: Gas6-mediated activation of Axl in melanoma cells.(A) Indicated melanoma cell lines were transiently transfected with control or Axl siRNA for 2 days prior to starving in quiescence media for 16h. Quiescent cells were then stimulated with indicated factors at the concentrations of 10 nM (EGF) and 200 ng/ml (Gas6) for 10 min followed by lysis in RIPA buffer for immunoblotting using the indicated antibodies. (B) Immunoblotting of indicated proteins extracted from quiescent melanoma cell lines treated with or without 200 ng/ml Gas6 for 10 min. NT: non treatment. Shown are representative blots of three experiments.

Fig 3: Stable knockdown of Axl limits migration and invasion of IgR3 cells.(A) Immunoblottings of stable anti-Axl shRNA expressing IgR3 monoclonal clones using indicated antibodies show selective downregulation of Axl. (B) Quantitative results of relative migratory speed of scramble and Axl shRNA clones. (C)Representative images of DAPI-stained cells invaded through MatriGel and retained on the bottom of transwell membrane. (D) Quantitative results of invaded cells. Shown are representative blots and pictographs of three experiments. Data are mean of ± SD of three independent experiments. Statistical analysis was performed using Student’s t-test. * P < 0.05, ** P< 0.01, NS: not significant.