Fig. 2

PAR1 exerts an oncogene, while PAR4 exerts a tumor suppressor in ESCC. A IHC detected the expression of PAR1 and PAR4 in ESCC tissues with or without (w/o) metastasis and representative photomicrographs of immunohistochemical staining were shown in B. C Western blot detected the expression of PAR1 and PAR4 at protein level. D Kaplan-Meier analysis of Disease Free Survival (DFS) related to the vWF expression in ESCC patients according to the data downloaded from TCGA database. The log-rank test was used to calculate the DFS rate for comparison between different groups. E Western blot detected the expression of PAR1 and PAR4 in HEEC, Kyse140 and TE-1 cells. F, G The effect of PAR1 or PAR4 KD on Kyse140 proliferation was analyzed by MTT assay. Cell viability was tested at 24th, 48th, 72th and 96th hour after transfection. H Wound healing assay detected migration ability of Kyse140 cells transfected with PAR1 or PAR4 shRNA. I 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 J. 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