FIG 9 .

Expression of the Grasp65-7A–GFP mutant inhibits AC morphogenesis. HF cell lines expressing either Grasp65wt-GFP or Grasp65-7A–GFP were plated on glass coverslips and allowed to grow until confluent prior to infection with HCMV AD 169 at an MOI of 1. Coverslips were collected 3 and 4 days p.i. and fixed with PFA. Coverslips were stained with antibodies to viral proteins pp65 (merge: blue), the envelope protein gM (merge: red), and DAPI (merge: gray). (A) The phospho-negative Grasp65-7A mutant inhibits AC formation. Panels show representative images of the AC in cells expressing Grasp65wt-GFP (top) or Grasp65-7A–GFP (bottom) at day 4 p.i. (B) Confocal microscopy was used to score the number of GFP-positive, HCMV-infected cells displaying a fully formed AC as shown in panel A. Percentages were calculated based on the total numbers of infected cells (Grasp65wt, n = 205; Grasp65-7A, n = 219) counted at each time point. The experiment was repeated 2 times, and the data shown are from a single experiment. Error bars show standard deviations.