FIG 9.

In vitro analysis of ASFVΔE66L. (A) ASFV or ASFVΔE66L infection induced the subversion of cell cycle in asynchronously growing cells. PAM cells were infected with ASFV or ASFVΔE66L at an MOI of 0.1. At 48 h, cells were collected and stained with PI for cell cycle analyses using flow cytometry. The data are from one of three experiments. The histograms were analyzed to determine the percentage of cells in each phase of the cell cycle. The results are shown as mean ± SD of three independent experiments; ns, not significant; **, P < 0.01. (B) Cellular protein synthesis during ASFV and ASFVΔE66L infection. Cultures of PAM cells (1 × 10⁶) were mock infected (Mock), infected with ASFV (0.1 MOI), or infected with ASFVΔE66L (0.1 MOI), and labeled at 24 h after infection with 3 μM puromycin for 1 h. Samples were analyzed using Western blot analysis. (C) Genes upregulated by infection with wild-type ASFV or the mutant ASFVΔE66L are displayed as a heat map. Colors represent the log₂ (normalized counts) values of the expression levels for the genes in the samples. (D) Quantitative real-time PCR analysis of selected RNAs (IFN-ALPHAOMEGA, IFNB1, TNF, MAP3K8, CCNL1, and CCNT2) in PAM cells. The results are shown as means ± SD of three independent experiments; *, P < 0.05; ***, P < 0.001.