Figure 7. PI3K is not required for FMDV entry and replication in BHK-21 cells.

(A,B) Wort moderately stimulated FMDV entry. Pretreated cells (40 μM Wort) were infected (MOI 25) for 1 h at 37 °C and processed for confocal microscopy with AF594-phalloidin (red), anti-FMDV (green), and DAPI (blue). (B) Quantitative analysis of the internalization of FMDV. The internalized FMDV were analyzed in 10 individual DMSO- or Wort-treated cells. Each experiment was performed in triplicate and the results were presented as the mean ± SD. (C–E) Wort enhanced FMDV infection. Pretreated cells (Wort) were infected (MOI 1) for 4 h at 37 °C and analyzed by RT-PCR (C), Western blot (D), and TCID50 assay (E). (F) Effect of Wort on virus entry and post-entry steps. Cells were treated with Wort 30 min before the infection (Pre) or treated 60 min after virus addition (Post) and maintained during the infection. Cells were then infected (MOI 1) for 4 h at 37 °C and processed for Western blot analysis. (G) PI3K downregulation moderately enhanced FMDV infection. (H) PI3K overexpression did not affect FMDV infection. 3D, FMDV 3D; β-actin, load control; SD, standard deviation; *P < 0.05; **P < 0.01.