Fig. 4.

P. aeruginosa infection upregulated autophagic degradation. (A) MH-S cells were infected with PAO1 at different times. Before infection, the cells were also treated with rapamycin (3 μM, 12 hours) and 3-MA (3 mM, 3 hours). (B) Western blotting of LC3 was performed. MH-S cells were treated with chloroquine (CQ; 40 μM, 6 hours) and then infected with PAO1 for 1 hour (MOI=10:1). Western blotting of LC3 was performed. (C) MH-S cells were transfected with GFP-LC3 plasmids for 24 hours, and then treated as in A. Western blotting of GFP was performed. GAPDH was used as a loading control in A–C. (D) MH-S cells were transfected with tandem GFP-RFP-LC3 plasmids for 24 hours. Then the cells were infected with PAO1 for 1 hour (MOI=10:1). Arrows indicate LC3 punctae, which could only be detected in RFP channel. (E) Puncta numbers in each cell was determined. The data are representative of 100 cells for each channel (one-way ANOVA; Tukey's post-hoc test, *P<0.05). Data are representative of three experiments with similar results.