Figure 2.

P. gingivalis-mediated upregulation of GARP enhances TGFβ activation through TLR4/MYD88 signaling. (A) The expression levels of indicated proteins in NE6-T and KYSE30 cells under different treatment conditions as determined by Western blot; (B) The expression level of active TGFβ secreted from NE6-T and KYSE30 cells in response to P. gingivalis was detected by ELISA; (C) The activity of a TGFβ-responsive Smad-binding element (SBE) luciferase reporter in GARP knockdown NE6-T and KYSE30 cells in response to P. gingivalis infection; (D) The expression levels of indicated proteins in NE6-T and KYSE30 cells under different treatment conditions as determined by Western blot. All experiments were independently repeated three times. (E) Representative images of IHC staining of TLR4 and MYD88 in tumor samples from ESCC patients with high- or low-levels of P. gingivalis. Scale bars, 200 µm and 100 µm, respectively. (F) Correlations between P. gingivalis and TLR4 protein, as well as between P. gingivalis and MYD88 protein. The quantitation data in (B and C) were presented as means ± SD from three independent experiment with a two-tailed Student’s t-test for statistical analysis. The Spearman nonparametric correlation test was employed to analyze the correlations among the ranked factors in (F) (*P < 0.05, **P < 0.01, and ***P < 0.001).