Fig. 5

Brequinar suppressed ASFV replication by activating ferroptosis. (A-C) PAMs were infected with ASFV (MOI = 1) at 37℃ for 2 h, and then treated with 100 μM brequinar in the absence or presence of uridine. After 24 h, the cells were washed thrice, and incubated with FerroOrange (A), Mito-FerroGreen (B), or Liperfluo (C) for 30 min. The fluorescence was visualized with confocal microscopy. (D, E) PAMs were infected with ASFV (MOI = 1) for 2 h, then 100 μM brequinar was added in the absence or presence of uridine (12.5, 25, 50 μM) or 50 μM uridine was added alone. The plate was further incubated for 24 h, and then the ASFV-B646L mRNA level was analyzed by qRT-PCR assay (D) and the p30 protein level was detected by western blotting assay (E). (F-I) PAMs were infected with ASFV (MOI = 1) for 2 h, then treated with cisplatin (10, 20, 40 μM) (F, G) or 100 μM brequinar in the absence or presence of ferrostatin (2.5, 5, 10 μM) (H, I). The plate was further incubated for 24 h. The B646L mRNA and p30 protein level was detected by western blotting assay and qRT-PCR assay, respectively. Statistical significance is denoted by *P < 0.05, **P < 0.01, and ***P < 0.001