FIG 3.

E4orf4 effects on DNA-PKcs autophosphorylation. (A) HCT116 cells were either mock infected or infected with the Ad mutant dl366*, lacking the E4 region, or with dl366*+E4orf4, lacking all E4 ORFs except E4orf4. Proteins were harvested at the indicated times p.i., and Western blot analysis was carried out with the specified antibodies. Samples from the 14- to 16-h and from the 24-h time points of this experiment were run on separate gels. (B) Protein bands in blots from two independent experiments performed as described above for panel A were quantified by densitometry, the levels of phosphorylated proteins were normalized to the levels of the corresponding total proteins, and the ratios between normalized phosphorylation in dl366*-infected cells (defined as 1) and parallel values in dl366*+E4orf4-infected cells are shown in a graph (n = 2). Error bars represent the standard errors. (C) HeLa cells were infected as described above for panel A and harvested for analysis at the indicated time points. All lanes shown are from the same exposed blot, but some lanes were removed from the middle.