Autophagy Contributes to Host Immunity and Protection against Zika Virus Infection via Type I IFN Signaling

Figure 5

Autophagy promoted ZIKV clearance in RAW264.7 cells. (a) Raw264.7 cells were transfected with ctrl siRNA or Atg7 and Atg5 siRNAs. qRT-PCR was performed for knockdown efficiency test. from triplicate. , . (b) Cells were infected with ZIKV for 1 hour, (). Before infection, the cells were also transfected with LC3-GFP and treated with Atg5 siRNA and Atg7 siRNA. Immunostaining was performed to detect LC3 puncta. (c) Puncta number in each cell was counted. Values are from 20 cells/sample. One-way ANOVA; Tukey’s post hoc test, . (d) RAW264.7 cells were infected with ZIKV for 1 hour (). Before infection, the cells were also treated with rapamycin and 3-MA; the rapamycin was used at (3 μM, 12 hours); 3-MA was used at (3 mM, 3 hours). The number of internalized virus per cell after 1 h infection was counted by PFU assay. After infection for 12 h, the unbound virus was washed away and fresh medium was added. The clearance assay was performed as counting the number of internalizing virus per cell by PFU assay. (e) Before infection, RAW264.7 cells were treated with negative control siRNA, Atg5 siRNA, and Atg7 siRNA. The number of internalized virus per cell was counted as above. Data were representative of three experiment results. . One-way ANOVA; Tukey’s post hoc test, , .

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