Figure 5
From: Porcine epidemic diarrhea virus activates PERK-ROS axis to benefit its replication in Vero E6 cells
Silencing of the ero1α gene reduced cytosolic ROS and viral replication in PEDV-infected Vero E6 cells. Both the ERO1α-silencing (shERO1α) Vero E6 cells and control cells (shNC) were infected with PEDV (MOI = 0.001) for 36 h. A The effects of ero1α knockdown on CHOP, ERO1α, and PEDV N expression were shown by Western blotting with β-actin used as loading control. B Ratios of ERO1α to β-actin. C Ratios of CHOP to β-actin. D Ratios of N to β-actin. E 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. F 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. The effects of ERO1α inhibition by shRNA on viral replication, shown as viral RNA copy numbers measured by RT-qPCR (G) and virus titer (H). The bar charts in B to D, F to H represent mean ± SD of three independent experiments. ns, not significant; *, P < 0.05; **, P < 0.01.