Fig. 2. Protein V is required for adenoviral replication in normal cells.

(A–C) Primary cells were infected with adenovirus at an MOI of 10 PFU/cell for various times. (A) Total DNA was extracted from infected cells, and 50 ng of total DNA was analyzed by qPCR. Plots represent as the copy number of the E4 gene per 50 ng of total DNA. (B) One-step growth curve kinetics of adenoviruses was analyzed in primary cell lines. (C) Proteins were extracted from infected primary cells at various times post-infection and 40 µg of total proteins were subjected to Western blot with the indicated antibodies. (D) Primary cells were infected with adenovirus at an MOI of 10, 50 or 100 PFU/cell for 1 and 48 hours. Crude proteins were extracted from infected cells, and 10 µg of proteins was subjected to Western blot. Ctrl: 1.0 × 10⁹ viral particles (VPs) of purified Ad5 were applied as a control for the pV expression. (E) Production level of infectious adenovirus in primary cell lines at 48 h.p.i. was measured by plaque assay. Results were reported as mean ± SD. N.S.; no significant difference