Fig 2.

Broad-spectrum anti-PRRSV assessment of hyperoside. MARC-145 cells were incubated with or without hyperoside (120 µM) for 2 h and then infected with various PRRSV-1 strains (GZ11-G1 and P073-3) at 37°C for 1 h. The cells and cellular supernatants were harvested at 24 hpi to evaluate PRRSV N protein expression via western blot analysis (A), and the progeny virus titer was determined via TCID₅₀ analysis (B). Moreover, the antiviral effects of hyperoside against PRRSV GZ11-G1 and P073-3 were also evaluated in PAMs. These cells were collected at 24 hpi to monitor N protein expression via western blot analysis (C). The cellular supernatants were also harvested to detect progeny virus production on the basis of the TCID₅₀ value (D). MARC-145 cells were pretreated with hyperoside (120 µM) or DMSO for 1 h and then incubated with different PRRSV-2 strains (CH-1a, JXA1, and NADC30-like; MOI 0.1) for 1 h. All cells and cellular supernatants were collected at 24 hpi to assess PRRSV N protein expression (E) and progeny virus titer (F). The antiviral effects of hyperoside against the PRRSV-2 strains CH-1a, JXA1, and NADC30-like were subsequently assessed in PAMs. PRRSV N protein expression was monitored at 24 hpi via western blot analysis (G), and the progeny virus titer was examined on the basis of the TCID₅₀ value (H). GAPDH served as the reference gene, and α-Tubulin served as the loading control. All the data are expressed as the means ± SDs and were subjected to Student’s t-tests. ***, P < 0.001, compared with DMSO-treated cells challenged with the same virus.