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Figure 2 | Veterinary Research

Figure 2

From: Tomatidine inhibits porcine epidemic diarrhea virus replication by targeting 3CL protease

Figure 2

Identification of anti-PEDV activity of tomatidine in Vero and IPEC-J2 cells. Vero cells were pretreated with the indicated concentrations of tomatidine (AD) for 1 h, and then infected with PEDV for 1 h at 37 °C. The cells were washed with PBS, then incubated in fresh medium containing tomatidine for 16 h. DMSO served as the treatment control. A Culture supernatants were collected at the indicated time points for viral titration. Results are expressed as TCID50. Titers from three independent experiments are shown as mean ± SD (error bars). B Western blot of N-protein in cells infected with PEDV and treated with the indicated concentrations of tomatidine or DMSO, at 16 hpi. C Relative PEDV N mRNA levels, determined by qRT-PCR, and expressed relative to that in DMSO-treated cells. The internal loading control was GAPDH. D Light microscope and IFA images of Vero cells infected with PEDV and treated with tomatidine, at 16 hpi. Viral N-protein is green. E Western blot of N-protein in cells infected with different PEDV genotypes and treated with indicated concentrations of tomatidine or DMSO, at 16 hpi. F, G IPEC-J2 cells were pretreated with tomatidine for 1 h, and infected with PEDV for 1 h at 37 °C. After incubation for a total of 24 h in fresh medium containing tomatidine, the culture supernatants were collected at the indicated time points for viral TCID50 titration, and the relative PEDV N mRNA levels in the cells were determined by qRT-PCR as above. H Viability of cells pretreated with the indicated concentrations of tomatidine and incubated for 24 h in medium containing tomatidine. The results are from one of three independent experiments. Error bars represent the SD. The asterisks in the figures indicate significant differences (*P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant).

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