Figure 8

PEDV traffics to lysosome via endosomes. A, B Vero cells and IPEC-J2 cells were transfected with siRab7 and siVPS39 twice, respectively, and then infected with PEDV strains at 24 h after the second transfection. The cells were collected at 6 hpi for qRT-PCR analysis. Ctrl means control. C–E The Vero cells and IPEC-J2 cells were pre-cooled at 4 °C for 15 min, incubated with PEDV strains at 4 °C for 1 h, shifted to 37 °C to initiate internalization. The non-internalized viral particles were removed by washing. 30 min after shifting, the cells were fixed and stained with anti-PEDV-S (red) and anti-EEA1 (green) primary antibodies (C). 40 min after shifting, the cells were fixed and stained with anti-PEDV-S (red) and anti-Rab7 (green) primary antibodies (D). 50 min after shifting, the cells were fixed and stained with anti-PEDV-S (red) and anti-LAMP1 (green) primary antibodies (E). Scale bars indicate 5 μm in C–E. *P < 0.05; **0.05 < P < 0.01; ***0.01 < P < 0.001; ****P < 0.001.