Fig. 2

RhoA knockdown enhances PRV infection. A Cell proliferation assay of PK-15 transfected with RhoA siRNA and control siRNA for 24 h. B Immunoblotting analysis of whole cell extracts from PK-15, NC, siRhoA-1 and siRhoA-2 PK-15 cells with antibodies against RhoA. β-actin served as a loading control. Gray value analysis of PRV-gB intensity by Image J software. C RT-qPCR analysis of RhoA mRNA in NC, siRhoA-1 and siRhoA-2 PK-15 cells. GAPDH served as a loading control. D RT-qPCR analysis of PRV gB and RhoA mRNA in NC, siRhoA-1 and siRhoA-2 PK-15 cells infected with PRV-QXX (MOI = 0.1) for 24 h. GAPDH served as a loading control. E Immunoblotting analysis of whole cell extracts from NC, siRhoA-1 and siRhoA-2 PK-15 cells infected with PRV-QXX (MOI = 1) for 24 h with antibody against PRV gB. GAPDH served as a loading control. Gray value analysis of PRV-gB intensity by Image J software. F PK-15 cells treated as in B and infected with PRV-QXX (MOI = 0.1 and 1), viruses were harvested with three freeze–thaw cycles, and the viral titer was determined by PFU assay. G Fluorescent microscopy and flow cytometry analysis of PRV-GFP (MOI = 0.01) proliferation in NC, siRhoA-1and siRhoA-2 PK-15 cells for 24 h. Scale bar, 200 μm. H NC, siRhoA-1 and siRhoA-2 PK-15 cells infected with PRV-QXX (MOI = 1) for 24 h, determination of PRV genome copy number based on PRV-gH. Data are shown as mean ± SD based on three independent experiments