FIG 3.

Dengue infection promotes global autophagy. To study autophagy, flux Huh7 cells were transfected with LC3 traffic light reporter plasmid p-mRFP-EGFP-LC3 and 16 h later were infected with the serotypes of dengue for 48 h. (A) Representative confocal images of mock-infected, EBSS-treated, and DENV-infected cells. The infected cells were stained with a serotype-specific dengue envelope antibody. (B) Quantification of the average number of red LC3 puncta per cell for the various serotypes at 48 h postinfection. About 20 cells were analyzed for each condition. (C) Heat map representing the transcript level of the indicated autophagy-specific genes in DENV-infected cells at the indicated time postinfection with respect to mock-infected cells. (D) Western blot analysis of p62 and LC3B I and II levels in mock- or DENV-infected Huh7 cells, either untreated or treated with EBSS (to induce autophagy) or bafilomycin (to suppress autophagy). PrM was used as an infection marker, and actin was used as an internal loading control. Postinfection (40 h), the cells were treated with EBSS for 1.5 h or bafilomycin-A for 8 h. Data are the mean ± SEM of three independent experiments. Statistical analysis was performed using one-way ANOVA. ****, P < 0.0001.