Fig. 3

PAR1 promoted the growth and metastasis of ESCC cell lines, while PAR4 had an inhibitory effect in vitro. A Fluorescence intensity represented the effects of PAR1/PAR4 on intracellular calcium mobilization in Kyse140 cells. B MTT assay detected the effect of PAR1 AP or PAR4 AP on Kyse140 proliferation. Cell viability was tested at 24th, 48th, 72th and 96th h. C The FACS detected PAR1 AP or PAR4 AP-induced ESCC cell apoptosis. Percentage of apoptotic cells compared to control was quantitated by mean fluorescence intensity. D Western blot analysis of apoptosis-related molecules in Kyse140 cells. β-actin was used as a loading control. E Wound healing assay detected migration ability of Kyse140 cells treated with PAR1 AP or PAR4 AP. F Transwell assay detected the migration ability of Kyse140. Cells which passed through the Transwell membranes were dissolved in methanol and quantified by microplate reader. OD value at 405nm wavelength was shown in G. Data are presented as mean ± SD from three independent experiments; comparison between two groups, *P < 0.05; **P < 0.01; ***P < 0.001. Scale bar: 200 μm in wound healing assay, 100 μm in Transwell assay