Fig. 1. NOP53 promotes the replication of HSV-1/F in Vero cells.

a,b Vero cells seeded in T-25 flask were transiently transfected with control (Ctrl) plasmid, plasmids to express Flag-tagged wt NOP53 (NOP) or truncated N4 (5 μg each) for 36 h. The cells were infected with HSV-1/F at 0.1 MOI, and intracellular viral yields a or extracellular viral yields b were determined 36 or 48 h postinfection (h.p.i.) by plaque assay. They are presented as log₁₀ PFU/ml. These experiments were performed two times with three replicates in each experiment. Values represent means with standard deviations (SD). *p ≤ 0.05; **p ≤ 0.01. c Vero cells were transfected with control plasmid, Flag-tagged NOP53, or Flag-tagged N4, and then infected with HSV-1/F at 0.1 MOI. Cell lysates prepared 36 or 48 h.p.i. were then analyzed by Western blotting using antibodies directed against HSV-ICP8 and Flag. Actin served as the loading control. d Vero cells seeded in T-25 flask were transfected with specific siRNA targeting NOP53 (siNOP) or negative siRNA (siNeg) (200 pmol each) for 72 h. The cells were then infected with HSV-1/F at 0.1 MOI, and viral yields were determined 36 or 48 h.p.i. by plaque assay. They are presented as log₁₀ PFU/ml. Values represent means of triplicates with SD. *p ≤ 0.05; **p ≤ 0.01. e Vero cells were transfected with siNOP or siNeg, followed by infection with HSV-1/F at 0.1 MOI. Cell lysates prepared 36 or 48 h.p.i. were then analyzed by Western blotting with the indicated antibodies