Skip to main content
. 2017 Sep 18;7:11759. doi: 10.1038/s41598-017-12173-1

Figure 4.

Figure 4

Involvement of PAR2 activation and subsequent phosphorylation of Akt and ERK1/2 in the P. gingivalis infection-induced microglial migration. (a) Representative images of migrated MG6 cells. The Boyden chamber assay was performed to evaluate cell migration after infection of MG6 cells with P. gingivalis in the presence or absence of control Ab (1 μg/ml), PAR2 neutralization antibody (SAM11; 1 μg/ml), control siRNA (50 nM) and PAR2 siRNA (50 nM). Microglial cells that migrated through a membrane were stained and counted after 12 h. Scale bar, 100 μm. (b) The quantitative analyses of the number of migrated cells. The results represent the mean ± SEM of three independent experiments. A one-way ANOVA with post hoc Tukey’s test; none vs. Pg: ***p = 0.0004, Pg + Con Ab vs. Pg + PAR2 Ab: †p = 0.0328, Pg + Con siRNA vs. Pg + PAR2 siRNA: ††p = 0.0036. (c) The immunoblots show p-Akt and total Akt after infection of MG6 cells with P. gingivalis in the presence or absence of PAR2 neutralization antibody (1 μg/ml) or PAR2 siRNA (50 nM) at 2 h. (d) The quantitative analyses of the p-Akt were shown. The results represent the mean ± SEM of three independent experiments. A one-way ANOVA with post hoc Tukey’s test; none vs. Pg: ***p = 0.0001, Pg + Con Ab vs. Pg + PAR2 Ab: ††p = 0.0027, Pg + Con siRNA vs. Pg + PAR2 siRNA: ††p = 0.0021. (e) The immunoblots show p-ERK1/2 and total ERK1/2 after infection of MG6 cells with P. gingivalis in the presence or absence of PAR2 neutralization antibody (1 μg/ml) or PAR2 siRNA (50 nM) at 30 min. (f) The quantitative analyses of the p-ERK were shown. The results represent the mean ± SEM of three independent experiments. A one-way ANOVA with post hoc Tukey’s test; none vs. Pg: ***p = 0.0001, Pg + Con Ab vs. Pg + PAR2 Ab: †p = 0.0103, Pg + Con siRNA vs. Pg + PAR2 siRNA: ††p = 0.0074. Full bots are presented in Supplementary Fig. S6.