Figure 6.

c-Met is an important adaptor in CagA-mediated autophagy pathway. (A,B) AGS cells were infected with Hp-WT or Hp-ΔcagA, and p-c-Met and c-Met were detected by western blot. CagA was immunoprecipitated from lysates. Immunoprecipitates (IP) were subjected to SDS-PAGE and immunoblot (IB) analysis with anti-p-c-Met (top) or anti–c-Met (bottom) antibodies. (C) Confocal microscopy showing AGS cells co-transfected with GFP-MAP1LC3B plasmid and c-Met siRNAs or control siRNA for 24 h, and then infected with Hp-WT or Hp-ΔcagA for 6 h. The percentages of cells with MAP1LC3B punctas are shown in the right graph with data being expressed as means ± SEM of three experiments (n ≥ 200 cells). (D) Western blot analysis of p-c-Met, MAP1LC3B-II conversion and β-actin in AGS cells transfected with c-Met siRNA or control siRNA and infected with Hp-WT or Hp-ΔcagA for 6 h. p-c-Met and MAP1LC3B-II band intensity was normalized to β-actin. (E,F) Flow cytometry showing MDC (upper panel) and AO (lower panel) staining of AGS cells transfected with c-Met siRNA or control siRNA and then infected with Hp-WT or Hp-ΔcagA for 6 h. (G) Western blot analysis of p-c-Met, MAP1LC3B-II conversion and β-actin in CagA-expressing AGS cells (AGS cells after transfecting the CagA expression plasmid, GFP-CagA) after transfected with c-Met siRNA or control siRNA and infected with H. pylori as described above. Experiments performed in triplicate showed consistent results. ^*P < 0.05, ^**P < 0.01.