Figure 3

Overexpression of H2BE facilitates PEDV replication. Marc-145 cells were transfected with H2BE-Flag or am empty vector. A High-level expression of H2BE was confirmed by Western blotting. Marc-145 cells were transfected with H2BE-Flag (2.5 μg) or the empty vector (2.5 μg) and then infected with PEDV at an MOI of 1 and harvested 12 h and 24 h later. B The PEDV N protein level was measured by Western blotting. C PEDV N mRNA was detected and analysed by qPCR. β-Actin was used as the internal control. D The intensity represents the PEDV N protein level normalized to that of β-actin. Marc-145 cells were transfected with H2BE-Flag (0, 2.5, 3 and 3.5 μg) and then infected with PEDV at an MOI of 1 and harvested 24 h later. E The PEDV N protein level was measured by Western blotting. F PEDV titres in the culture supernatants were determined using the TCID₅₀ method. The results are representative of three independent experiments. The data are presented as the mean ± SD, n = 3, (*P < 0.05; **P < 0.01). G Marc-145 cells were transfected with H2BE-Flag or the empty vector and then infected with PEDV at an MOI of 1.0. Cells were fixed and incubated with mouse anti-PEDV Nsp9 polyclonal sera (1:500) 24 h post-infection. Immunofluorescence assays were used to further observe intracellular propagation of PEDV. H Analysis of H2BE protein fluorescence intensity.