ATM and ATR inhibit Ad replication whereas E4orf4 enhances it.

(A) ATM-deficient A-T cells, reconstituted with WT ATM or with an empty vector, were infected with the Ad mutants dl366* and dl366*+E4orf4 (30 ffu/cell), or were mock infected. An ATR inhibitor (ATRi) was added to parallel cell samples as described in the methods section. The cells were visualized 24 hrs post infection using a Zeiss Axioskop at a magnification of 100. Representative pictures of two independent experiments are shown. The percent of area covered with cells out of the total field area (ce) was determined using Image-Pro Premier 9.1 and the percent of area covered with cell clusters larger than 6000 pixels2 out of total cell area (cl), was calculated using FIJI. (B) Viruses from two independent experiments performed as described in A were collected from infected cells 24 hrs post infection and their titer was determined. Data is displayed in box plots as previously described [72] and black dots represent outliers. Statistical significance was calculated using unpaired t-test and p values of significant differences between dl366* and dl366*+E4orf4 viruses are shown.