Figure 3.

Knockdown of endogenous ATP1A1 expression suppresses PEDV infection. (A,B) Knockdown of endogenous ATP1A1 significantly suppresses PEDV RNA abundance. Transfected with siRNAs against ATP1A1 or siRNA-NC for 24 h, cells were collected at 12, 24 and 48 h after PEDV (0.1 MOI) infection of target cells for quantitative PCR assay. Data represent means ± SD from three independent experiments. ***, p < 0.001. (C,D) Knockdown of endogenous ATP1A1 reduces in PEDV N protein expression. After knocking down with the expression of ATP1A1 for 24 h, target cells were infected with PEDV (CV777-G1 or GDgh-G2) at 0.1 MOI and collected and lysed at different time points for Western blot analysis. (E,F) Knockdown of endogenous ATP1A1 suppresses PEDV viral titers. PEDV at 0.1 MOI infected cells with low ATP1A1 expression for 24 h, and cell supernatants were extracted for TCID₅₀ assay. Data represent means ± SD from three independent experiments. *, p < 0.05. (G,H) Knockdown of endogenous ATP1A1 reduces PEDV infection. Cells were transfected with siRNAs for 24 h, and then PEDV (0.1 MOI) infected for 24 h. The cells were stained with anti-ATP1A1 mAb (red) and anti-PEDV-N mAb (green). Staining of cell nuclei using DAPI (blue) staining solution. The processed samples were photographed and analyzed using fluorescence microscopy. Scale bars, 200 μm.