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Figure 4 | Veterinary Research

Figure 4

From: Xanthohumol inhibits PRRSV proliferation and alleviates oxidative stress induced by PRRSV via the Nrf2–HMOX1 axis

Figure 4

Xn inhibits PRRSV adsorption and internalization in Marc-145 cells. A Adsorption assay. Cells were incubated with a mixture of Xn or DMSO and PRRSV for 1 h at 4 °C and then harvested for qRT-PCR. B Internalization assay. Cells were treated with CHX (10 μg/mL) for 12 h first, then incubated with PRRSV (1 MOI) for 1 h at 4 °C, washed, and finally incubated with 10 µM Xn or DMSO for another 1 h at 37 °C. The levels of PRRSV ORF7 mRNA were detected by qRT-PCR, and the visible virions in cells were detected using IFA confocal laser-scanning microscopy. Scale bar 10 µm. C Cells were infected with PRRSV (1 MOI) for 6 h, and culture medium was replaced with 2% DMEM containing Xn (10 µM) or DMSO. At 7, 8, 9, and 10 hpi, the PRRSV ORF7 levels were measured using qRT-PCR. D Cells were infected with PRRSV (0.1 MOI) for 18 h, then washed and incubated with fresh DMEM containing 10 µM Xn or DMSO. Supernatants were harvested, and released virus was titred by plaque assay. qRT-PCR results are presented relative to the reference control (mock infected cells). GAPDH was used as the internal loading control. All assays were repeated at least three times, with each experiment performed in triplicate. Bars represent the means ± SEMs from three independent experiments. **P < 0.01; *P < 0.05 compared with the DMSO-treated group

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