FIG. 10.

ATR and ATRIP are not targets of ICP0. (A and B) HFF-1 cells were infected twice with lentiviruses expressing either shGFP or shATRIP2. Cells were then infected with wild-type HSV-1 (in1863) (A) or ICP0-null (dl1403/CMVlacZ) (B) at dilutions suitable for plaque assays. The relative probability of plaque formation on shATRIP2 cells was determined with respect to the same dilution of shGFP cells. The values represent the averages from three independent experiments, and the error bars represent the standard errors of the means. (C) HeLa cells were infected with d106, which expresses only ICP0, at an MOI of 1 or 10, and samples were harvested at 6 and 24 h postinfection as described in Materials and Methods and separated by SDS-PAGE. Western blotting was performed for ATR N19, rATRIP Upstate, Chk1, and PML. ICP0 served as an infection control, and Ku70 served as a loading control. (D) Vero cells were transfected with Myc-ATRIP, Flag-ATR, and ICP0 as indicated. ATRIP immunoprecipitation was performed from cell lysates with Myc antibody, and the products were separated by SDS-PAGE and blotted with Myc, ATR N19, and ICP0.