Fig 2.

Tunicamycin inhibited viral attachment, internalization, and entry of SADS-CoV. (A) Vero cells were pretreated with tunicamycin for 24 h and then infected with SADS-CoV (MOI = 10) for 1 h on ice. Cells were then washed with cold PBS three times, and viral attachment on the cell surface was detected by RT-qPCR. (B) After viral attachment for 1 h on ice, cells were cultured at 37°C for an additional 1 h, and viral internalization was detected by RT-qPCR. For viral entry, Vero and Huh-7 cells were pretreated with tunicamycin for 24 h and then infected with SADS-CoV (MOI = 1) for 6 h. Cells were lysed, and virus detection was performed using qRT-PCR (C). Additionally, cells were fixed and stained with anti-SADS-CoV NP antibodies for analysis by IFA (D). All data are shown as means ± standard errors of the means (n = 3 biological replicates). Scale bars: 300 µm.