HCMV UL97 interacts with 14-3-3 β, ε, η, γ, and θ.A, U-2 OS cells were transfected with an expression vector for HA-tagged UL97 or an empty vector (−) together with expression vectors for one of each of the seven isoforms of the Flag-tagged 14-3-3 proteins. After 48 h, lysates (Input) were harvested, subjected to immunoprecipitation (IP) with anti-HA antibody (IP: anti-HA) and analyzed by Western blot with the indicated antibodies. B, hTERT-immortalized human foreskin fibroblasts (HFF2/T) were mock infected (M) or infected with HCMV (strain AD169) at an MOI of 3 in the presence of 10% FBS. Lysates prepared at the indicated hours post-infection (hpi) were analyzed by Western blotting with the indicated antibodies. C, lysates (Input) from panel (B) (48 hpi) were subjected to IP with normal rabbit IgG as a control and anti-14-3-3 ε, θ, and UL97 antibodies (IP: anti-C, ε, θ, 97) and analyzed by Western blot with the indicated antibodies. D, IPs as in panel (C) from 72 hpi lysates. For all panels, representative images from one of three independent biological replicates are shown. FBS, fetal bovine serum; MOI, multiplicity of infection.