Fig 2.

Effect of Rab5 on live P. aeruginosa invasion in macrophages. (A) J774-Eclone cells expressing GFP were incubated with live (L) or heat-inactivated, dead (D) strain PAO1 P. aeruginosa at a ratio of 200:1 for 5, 15, or 30 min. The phagocytic index was determined and is expressed as percent phagocytosis of heat-inactivated bacteria internalized in GFP control cells at 5 min. (B) J774-Eclone cells expressing GFP or Rab5:WT, Rab5:Q79L (QL), or Rab5:S34N (SN) were incubated with live or heat-inactivated P. aeruginosa at a ratio of 200:1 for 15 min at 37°C. After incubation, the phagocytic index was determined and is expressed as percent phagocytosis of heat-inactivated bacteria by GFP control cells at 15 min. (C) J774-Eclone cells alone (control) or cells transfected with a scramble RNAi sequence or RNAi sequences against Rab5 isoforms were incubated with heat-inactivated P. aeruginosa at a ratio of 200:1 for 30 min at 37°C. After incubation, the phagocytic index was determined and is expressed as the percent phagocytosis of control cells. Data represent the mean ± SEM from three independent experiments. Asterisks represent statistically significant differences from control group values (*, P < 0.05).