FIG 1.

ASFV-ΔH240R infection activates the NF-κB signaling to promote pro-IL-1β transcription. (A) pro-IL-1β transcription was upregulated in PAMs infected with ASFV-ΔH240R compared with ASFV-WT. PAMs were infected with equal genome copies of ASFV-ΔH240R and ASFV-WT for 12 h or 24 h, and pro-IL-1β transcription was examined by RT-qPCR using primers against pro-IL-1β and GAPDH. (B) The phosphorylation levels of IκBα and p65 were increased in the PAMs infected with ASFV-ΔH240R compared with ASFV-WT. PAMs were infected with equal genome copies of ASFV-ΔH240R and ASFV-WT for 12 or 24 h, and the phosphorylation levels of IκBα and p65 were analyzed by Western blotting. The protein band intensities were analyzed by Image Studio software. (C) pro-IL-1β transcription was decreased in the inhibitor-treated PAMs induced by ASFV-ΔH240R infection. BAY11-7082, an inhibitor of the NF-κB signaling pathway, was added to the PAMs infected with ASFV-ΔH240R or ASFV-WT, and the transcription of pro-IL-1β was analyzed by RT-qPCR at 12 or 24 hpi. (D) The phosphorylation levels of IκBα and p65 were decreased in the inhibitor-treated PAMs induced by ASFV-ΔH240R infection. BAY11-7082 was added to the PAMs infected with ASFV-ΔH240R or ASFV-WT, and the phosphorylation levels of IκBα and p65 were analyzed by Western blotting at 12 or 24 hpi. Data are from three independent experiments. The error bars denote standard errors of the means. The results of densitometry analysis to quantify the ratio of p-p65 or p-IκBα to GAPDH are shown at the bottom. The significance of differences between groups (n = 3) was determined using Student’s t test (***, P < 0.001; ns, not significant).