c-Abl kinase affects PDGF induced A375 migration.

<p>(A) a, the serum-starved cell suspension, with or without 10 µM STI571 (c-Abl kinase inhibitor), was seeded onto 0.8 μm diameter upper transwell chamber, which was coated with fibronectin. Free-serum medium with 50 ng/ml PDGF was put into the lower chamber to induce cell migration. b, migrations were determined by counting cells in randomly selected five microscope field per well. Bars represent mean ± S.D. (B) the effects of different concentration of STI571 on cell migration. The migration experiment of A375 cells out of agarose drop explants treated with or without STI571 was conducted. After 24 h, the distant of migration was measured using inverted microscope fitted with a rule in eyepiece. a, Cell migration rate was evaluated by the distance of the leading edge of migrating cells from the edge of agarose droplet. b, The distance of cell migration was measured, the extent of cell migration within the drop = [(total area/drop area) ×100] –100. (C) A375 cells were transfected with WT c-Abl expression vector or specific siRNA targeting c-Abl. The migration experiment of A375 cell or tansfected cells was performed in transwell plates (b was the statistic value of a). (D) A375 cells or cells transfected with WT c-Abl or c-Abl specific siRNA were lysed (For the transfected cells, cells were lysed after 24 h transfection). Lysates were blotted with c-Abl and actin antibodies. (E) a. A375 cells or the cells transfected with WT c-Abl or c-Abl specific siRNA were plated into 24-well plated and allowed to form a confluent monolayer. After overnight serum starvation, the cell monolayer was “wounded” with a pipette tip. The cells were incubated with DMEM containing 20 ng/ml PDGF and then imaged. b. Cell migration was determined as shrinkage of an average gap area. Bars represent mean ± S.D of three independent experiments. **, p<0.01, ***, p<0.001 with respect to control.