Figure 2.

Deletion of UL24 promotes NF-κB activation. (A) Diagram of the PRV UL24 gene, with the sgRNA1 and sgRNA2 incision positions. (B) The UL24 knockout virus was identified by PCR. Several clones were randomly chosen and cultured in Vero cells, and the virus genome was extracted and identified by PCR with the primers. (C) UL24 knockout was confirmed by DNA sequencing; 290 bases were deleted from 96~385 bp of the UL24 gene. The deletion region is shown as a dotted line. (D) UL24 knockout was confirmed by Western blot. (E) The HEK293 cells were inoculated with WT PRV or UL24 knockout PRV at a multiple of infection (MOI) of 0.1. The virus was collected at 4, 8, 16, and 24 h post infection. qPCR was used to quantify the copy numbers of viral DNA. (F) HEK293T cells were transfected with an NF-κB-Luc reporter plasmid. Twelve hours later, the cells were infected with HeN1 PRV or UL24 knockout PRV (MOI = 10), and luciferase activity was measured at 2 h, 4 h, 6 h, and 8 h post infection. The experiments were performed three times, and a representative result is shown. * indicates p < 0.05.