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. 2023 Feb 16;24(4):4000. doi: 10.3390/ijms24044000

Figure 7.

Figure 7

Knockdown of ATP1A1 affects the attachment of PEDV. (A) Knockdown of ATP1A1 significantly suppresses attached PEDV. Transfection with siRNAs against ATP1A1 and siRNA-NC for 24 h. In the attachment assay, PEDV at 1.0 MOI infected target cells at 4 °C for 2 h and then cells were washed with PBS and were collected to detect viral RNA abundance. In the internalization assay, PEDV at 1.0 MOI infected target cells at 4 °C for 2 h and then cells were washed with PBS and transferred to 37 °C for 1 h incubation to complete virus internalization. After washing the cells with PBS, the cells were treated sequentially with 0.05% trypsin and 0.5 mg/mL proteinase K to remove the uninternalized virus particles, and the cells were collected for viral RNA detection. Data represent means ± SD from three independent experiments. ***, p < 0.001. (B,C) S1 number of positive cells bound to ATP1A1. Incubating IPEC-J2 and Vero-E6 on ice with PEDV at 1.0 MOI for 1 h to synchronize infection, then transferring to 37 °C incubator for the indicated time. Cells were collected after washing with PBS at the appropriate time points. Cells were then stained using rabbit anti-ATP1A1 mAb and mouse anti-S1 mAb, followed by CoraLite488-conjugated Goat Anti-Rabbit IgG and CoraLite594-conjugated Goat Anti-Mouse IgG. Data analysis using flow cytometry.