Figure 7

Scavenging of ROS by N-acetyl-L-cysteine reduced CHOP and ERO1α expression, decreased cytosolic ROS, and limited viral replication in PEDV-infected Vero E6 cells. Vero E6 cells were infected with PEDV (MOI = 0.001) in the presence or absence of N-acetyl-L-cysteine (NAC, 10 mM) for 36 h. A Cells collected at 36 hpi were measured for changes in cytosolic ROS level by flow cytometry after incubation with 10 μM DCFH-DA for 30 min at 37 °C. B The changes in cytosolic ROS levels were evaluated according to the mean value of DCFH-DA fluorescence intensity and expressed as fold changes between mock- and treated-cells. C The effects of ROS scavenging on CHOP, ERO1α, and PEDV N expression were shown by Western blotting with β-actin used as loading control. D Ratios of CHOP to β-actin. E Ratios of ERO1α to β-actin. F Ratios of N to β-actin. The effects of ROS inhibition on viral replication, shown as viral RNA copy numbers measured by RT-qPCR (G) and virus titer (H). The bar charts in B, D to H represent mean ± SD of three independent experiments. *, P < 0.05; **, P < 0.01.