Production of wt-Ad3GFP and mu-Ad3GFP.

A) 293 cells were infected at indicated MOIs (vp/cell) for 2 hours. Medium samples were collected at indicated time points. Viruses were titrated on 293 cells based on GFP-expressing units (“Infectious Units”). N = 3. B) Subconfluent T84 cells were infected and progeny virus production was measured as described in A). C) Comparison of penton, fiber, and hexon expression. HeLa cells infected with wt-Ad3GFP or mu-Ad3GFP were collected at 36 hours post-infection and total cell lysates (without prior ultracentrifugation) were subjected to Western blot with anti-PtDd antibodies (recognize Ad3 penton base and fiber), anti-Ad antibodies (recognize Ad3 hexon, penton base, and fiber) and anti-actin (loading control) antibodies.