Stable expression of PACS-1Admut causes gB but not gH to be mislocalized during HCMV infection. (A) Stable U373-PACS-1, PACS-1Admut, or control cell lines were treated with or without 1 μg of doxycycline/ml for 2 days. Protein expression was assessed by Western blotting with PACS-1-specific antisera. (B) Stable U373 cell lines expressing empty vector controls (a to c and j to l), wild-type PACS-1 (d to f and m to o), or PACS-1Admut (g to i and p to r) were infected with HCMV at an MOI of 3 for 3 days in the presence of 1 μg of doxycycline/ml. Cells were fixed and developed with antiserum specific for HCMV gB (R2448; a, d, g, j, m, and p), together with antiserum specific for γ-adaptin (100/3; b, e, and h) or HCMV gH (14-4b; k, n, and q). HCMV gB staining is shown in green, and γ-adaptin or HCMV gH staining is shown in red.