Fig. 4

Overexpression of RhoA inhibits PRV replication. A Immunoblotting analysis of whole cell extracts from PK-15 cells expressing vector or FLAG-RhoA with antibody against RhoA. GAPDH served as a loading control. B RT-qPCR analysis of PRV gB and RhoA mRNA level in PK-15 cells expressing vector or FLAG-RhoA infected with PRV-QXX (MOI = 0.1) for 24 h. GAPDH served as a loading control. C Immunoblotting analysis of whole cell extracts from PK-15 cells expressing vector or FLAG-RhoA infected with PRV-QXX (MOI = 1) for 24 h with antibodies against PRV gB and FLAG. GAPDH served as a loading control. D Gray value analysis of C using Image J software. E Fluorescent microscopy and flow cytometry analysis of PRV-GFP (MOI = 0.01) proliferation in PK15 cells expressing vector or FLAG-RhoA for 36 h. Scale bar, 200 μm. F PFU assay in vector or FLAG-RhoA PK-15 cells infected with PRV-QXX (MOI = 0.1 or 1) for 24 h. G PK-15 cells treated as in B and 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