Figure 2

GBP1 overexpression inhibits nuclear delivery of PRV virions, which correlates with a disruption of the morphological actin filaments. PK-15 cells were stably transfected with pCI-GBP1 vector or the control vector pCI-neo, to establish PK-15^GBP1 and PK-15^PCI cells selected by G418. A Representative Western blots of HA-GBP1 protein in described cells. B, C PK-15, PK-15^GBP1, and PK-15^PCI cells were infected with PRV (1 MOI) for 24 h. Relative change in amount of the PRV genome copy number detected by q-PCR B Representative Western blots of HA-GBP1, gC and GAPDH. D Quantification of panel C. E–G PK-15 cells were harvested at the appointed times for detecting the mRNA levels of viral genes (immediate-early gene: IE180 at 3 hpi, early genes: EP0, UL54, UL48 at 4 hpi, and late genes: gD, gB, gC at 12 hpi). Relative change in the amount of viral gene mRNA. H These Cells were incubated with 10 MOI PRV for 1 h at 4 ℃, followed by three washes with cold PBS (for the adhesion group), and an additional hour of incubation at 37 °C (for the entry group). Next, total DNA was exacted and subjected to q-PCR. Relative change in amount of viral genome copy number. I Cells were incubated with 10 MOI PRV at 37 ℃ for 2 h, then the nuclear fraction was isolated to extract DNA for detecting the PRV genome copy number. Relative change in amount of viral genome copy number in the cell nucleus. J Representative immunofluorescent stain of DAPI (blue), HA-GBP1 (cherry), and F-actin (green). Scale bar, 20 μm. ** P < 0.01, versus the wild-type PK-15 cells (WT-PK).