Fig. 6

nSMase2/MAPK/NF-κB mediated PAR4 activation-induced tumor inhibitory effect, and FAK/PI3K/AKT/STAT3/NF-κB mediated PAR1 activation-induced tumor promoting effect in ESCC. A Kyse140 cells were lysed and immunoprecipitated with antibodies against IgG control, PAR1 or PAR4 and then immunoblotted with antibodies against nSMase2, PAR1 or PAR4. B Western blot detected the expression of p-p38 MAPK. C Kyse140 cells were pre-incubated with 30µM 3-OMS, followed by the addition of either PAR1-AP or PAR4 AP to trigger the phosphorylation of IKKβ. The IKKβ phosphorylation was detected by FCM. D Kyse140 cells were pre-treated with PAR1 AP or PAR4 AP. FAK phosphorylation was detected by FCM. E Western blot detected the expression of PI3K, p-Akt and Akt. F MTT detected the effect of nSMase inhibitor on Kyse140 proliferation. G Western blot verified the knockdown efficiency of shAKT plasmids. H Immunofluorescence staining and confocal microscopy detected p-STAT3 and p-NF-κB expression in control and AKT-knockdown Kyse140 cells. Data are presented as mean ± SD from three independent experiments; comparison between two groups, *P < 0.05; **P < 0.01; ***P < 0.001