Fig. 3

RhoA agonist inhibits PRV replication. A PK-15 cells were treated with Narciclasine (0–10 nmol/L) for 24 h. CCK-8 assays were then performed to determine the cell viability (%). B PK-15 cells were pretreated with Narciclasine at indicated concentrations for 4 h and infected with PRV-QXX (MOI = 0.1) for 24 h, expression analysis of PRV gB and TK mRNA levels using RT-qPCR. GAPDH served as a loading control. C Immunoblotting analysis of PK-15 cells treated as in B and infected with PRV-QXX (MOI = 1) for 24 h with the anti-PRV-gB antibody. GAPDH served as a loading control. D Gray value analysis of PRV gB using Image J software. E After treatment as described in B, the cells were fixed and stained with antibody of PRV gB (in green) and the nuclei were stained with DAPI. The fluorescence was measured by Zeiss LSM 800 microscope. Scale bar, 10 μm. F PK-15 cells treated as in B, viruses were harvested with three freeze–thaw cycles, and the viral titer was determined with PFU assays. G PK-15 cells treated as in B and infected with PRV-QXX (MOI = 0.1) for 24 h, determination of PRV genome copy number based on PRV-gH. All the data are shown as mean ± SD based on three independent experiments