PI3K-AKT signaling is downstream of EGFR in M. hyorhinis -infected MGC803 cells. (A) AG1478 or wortmannin pre-treatment abolishes M. hyorhinis-induced AKT phosphorylation. Prior to M. hyorhinis infection, cells were pretreated with 5 μM AG1478 or 2 μM wortmannin for 1 hour. After that, Western blot were performed. (B) Validation of silencing efficiency of EGFR. Cells were transiently transfected with 50 nM EGFR-specific siRNAs or a control siRNA. 48 hours after transfection, cell lysates were subjected to Western blot. (C) Knock down of EGFR abolishes M. hyorhinis-induced AKT phosphorylation. Cells were transiently transfected with 50 nM specific siRNA targeting EGFR. 48 hours after transfection, cells were infected with M. hyorhinis for 24 hours, followed by Western blot. Optical densities of protein bands were quantified by Image J software and relative expression levels of indicated protein to loading control were shown in graph. Values represented the mean ± SD from three independent experiments.