Fig. 5.

EGFL9 specifically activates cMET regulated signaling pathways. a Monitoring the receptor tyrosine kinase activation using a phospho-receptor tyrosine kinase array. Two isogenic cell lines, HMLE/LacZ and HMLE/EGFL9, were analyzed for signaling activation of a set of 49 receptor tyrosine kinases. b Quantitation of activation of cMET, EGFR, and IGR-IR in HMLE/EGFL9 cells in comparison with HMLE/LacZ. c Western blotting analysis of ectopic expression of EGFL9 in HMLE cells (left panel) and knockdown of EGFL9 in SUM159 cells. Phosphorylation and total cMET and cMET downstream targets (EGFR, FAK, AKT, and ERK) were examined. d Western blotting of cMET and its downstream targets in HMLE/EGFL9 cells after treatment with various concentrations of JNJ38877605 for 24 h. The antibodies used for the western blotting analysis were listed in the panels. e The treatment with JNJ38877605 leads to a significant decrease of cell migration (top panel, ***P < 0.001), invasion (bottom panel, ***P < 0.001) in HMLE/EGFL9, but not in HMLE/LacZ cells. Each bar represents the mean ± SEM (standard error of the mean). For three independent experiments, P value was determined by unpaired two-tailed t-test. f Western blotting of cMET and its downstream targets in SUM159 cells after treatment with various concentrations of JNJ38877605 for 24 h. The antibodies used for the western blotting analysis were listed in the panels