Figure 2
c-Met is activated by HGF in medulloblastoma cells and promotes wound closure. (A) IB of DAOY and UW228 lysates of cells after HGF stimulation. Antibodies used as indicated to the right of the panels. B) IB analysis of PHA-665752 and ARQ 197 (1 μM, 2 h pretreatment) effects on HGF-induced (50 ng/mL, 10 min) c-Met and ERK pathway activation (anti-p-c-Met and anti-pERK, respectively). (C) IB analysis as in B of the effect of prolonged (24 h) pretreatment with ARQ197 inhibitor on HGF-induced c-Met and ERK activation in UW228 cells. (D) IB analysis of HGF-induced activation of JNK and ERK in DAOY cells (20 ng/mL HGF −/+ 250 nM PHA-665752). (E) Schematic showing how the area covered by the cells in the Oris migration assay was determined. Light grey: cell monolayer at T0h. White: gap after removal of the plug. Dark grey: area covered by cells at T10h. (F) Quantification (means + S.D.) of Oris migration assays using DAOY or UW228 cells in HGF-treated (20 ng/mL) medium containing 0% FCS and pharmacological inhibitors (PHA-665752 and ARQ 197, both at 125 nM). (G) As F) but progression of gap closure shown for 0–10 h only.