Figure 2

Pharmacological inhibition of autophagy restrained DHAV-1 proliferation and viral release. A Pharmacology did not affect cell viability. After DEFs were treated with 100 nmol/L RAPA, 50 μmol/L CQ or the solvent DMSO, as described in Materials and methods, the cell viability of each group was assessed by the WST-1 assay. The optical density of sample wells was determined at 450 nm (OD₄₅₀) and normalized to the optical density of the blank well. Data shown represent the mean ± SD of three independent experiments. “ns” indicates no significant difference, P > 0.05. B–D After pretreatment with CQ (50 μmol/L) or RAPA (100 nmol/L) or no pretreatment, DEFs were infected with DHAV-1, and the number of extracellular and intracellular DHAV-1 viral copies was detected by RT–qPCR. All data were derived from three separate sets of experiments. E, F DEFs were treated with 50 μmol/L CQ before DHAV-1 infection, the cells were observed, and cytotoxic effects were recorded. Viral titres were calculated and are expressed as TCID₅₀ per mL. Differences between two groups were analysed using Student’s t test and are presented as P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***).