Fig. 5

Actin cytoskeleton polymerization contributes to PRV replication in vitro. A PK-15 cells were treated with different concentrations of Narciclasine for 4 h, infected with PRV-QXX (MOI = 0.1) for 24 h with the above concentrations of compound. The cells were fixed with 4% PFA, stained with anti-gB antibody for viral glycoprotein gB, phalloidin for F-actin and DAPI for nucleus. Fluorescence was analyzed by confocal microscopy. Scale bar, 10 μm. B PK-15 cells treated with cytochalasin D (0–500 nmol/L) for 24 h, CCK-8 assays were then performed to determine the cell viability (%). C RT-qPCR analysis of PRV gB and TK mRNA expression in cytochalasin D (0–500 nmol/L) treated cells infected with PRV-QXX (MOI = 0.1) for 24 h. GAPDH served as a loading control. D PK-15 cells treated as in C and infected with PRV-QXX (MOI = 0.1 or 1) for 24 h, then viruses were harvested with three freeze–thaw cycles, and the viral titer was determined with PFU assays. E Fluorescence microscopy and flow cytometry analysis of PK-15 cells infected with PRV-GFP (MOI = 0.01) for 24 h after treatment with cytochalasin D (0–500 nmol/L). Scale bar, 100 μm. Data are shown as mean ± SD based on three independent experiments