FIG 3.

Analysis of viral protein synthesis, cellular protein processing, and cellular innate immune induction during infection. (A) Primary swine kidney (PK) cells were infected with A12-WT and A12-P1 deopt at an MOI of 10 for 6.5 h. At the indicated times, cytoplasmic extracts were prepared and analyzed by Western blotting using rabbit anti-eIF4G polyclonal Ab, rabbit anti-VP1 polyclonal Ab, mouse MAb 5D8C7 (anti-L^pro), MAb F19 (305) (anti-3D), and mouse anti-tubulin-α MAb (Ab-2 MS-581). CP, eIF4G cleavage products. (B) Expression of IFN-β, TNF-α, RANTES, Mx1, IL-1β, and IRF7 mRNAs was measured by real-time RT-PCR in PK cells infected with A12-WT and A12-P1 deopt FMDV at an MOI of 2 for the indicated times. Porcine GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as an internal control. Results are expressed as fold increase (antilog ΔΔC_T) of gene expression for virus-infected versus mock-infected cells (reported values display results of one out of three representative experiments with similar results). (C) Porcine IFN-α was detected by ELISA in the supernatants of PK cells infected with A12-WT and A12-P1 deopt FMDV at an MOI of 2. The values are presented as the mean ± standard deviation (SD) from three independent determinations.