Fig. 5
PAR2 activation mediates enhanced migration of LX-2 cells in a Met-, PDGFR-, Src- kinase-, p42/p44 MAPK- and MMP-dependent manner. a LX-2 cells (wt, shPAR2 or shCo as indicated) were serum-starved for 17 h and treated with PAR2-AP (10 μM), or trypsin (10 nM) for 6 h. Cells had migrated through the collagen barrier and the pores of the polycarbonate membrane were fixed, stained and quantified by microscopic counting. Bars represent the mean values ± S.D. of octuplicates obtained in one experiment, which is representative for three independent assays. **P-value < 0.05 versus non stimulated control. b Serum-starved LX-2-wt cells were stimulated with PAR2-AP (10 μM) for 4 h and analysis of plasma membrane filopodial structures was performed using scanning electron microscopy as described in the method section. (a) Cells with filopodia were quantified in a blinded manner from 100 individual cells per each group and from three experimental preparations. (b) enlarged inset. The picture shows a single LX-2-wt cell with typical filopodial spike pattern after stimulation with PAR2-AP. c Serum-starved LX-2-wt cells were preincubated for 1 h with vehicle, SU 11274 (10 μM), PHA 665752 (0.1 μM), AG 1296 (1.0 μM), SL 372 (5.0 μM), PD 98059 (10 μM), U 0126 (10 μM), PP2 (5.0 μM), or GM-6001 (1.0 μM). Cell migration in response to PAR2-AP (10 μM) was analysed after 6 h as described under a. Representative results from three independent experiments are shown. *P-value < 0.05 versus non stimulated control, **P-value < 0.05 versus stimulated with PAR2-AP and versus non stimulated control