Fig. 3.

Specific interaction between endogenous emerin and pUL50 in transfected and HCMV-infected cells. A–F, FLAG-/HA-tagged HCMV proteins, pUL50 deletion mutants, or pcDNA3.1 (vector) were transiently expressed in 293T cells as indicated. Cells were lysed at 2 days post-transfection, and this was followed by immunoprecipitation of the FLAG-tagged viral proteins and the HA-tagged pUL50 deletion mutants using mouse mAb-FLAG (A) and rabbit pAb-HA (C and E), respectively. A mouse Fab fragment was used as a control for specificity (C and E, lane 1). Coimmunoprecipitates and expression control samples were subjected to Wb analysis using tag-specific antibodies or mAb-emerin. G, H, HFFs were infected with HCMV strain AD169 at an MOI of 0.1 or remained uninfected (mock). At 3 dpi, cells were lysed and used for CoIP analysis with mAb-emerin. Detection of coimmunoprecipitates and of expression controls was performed by means of Western blotting using mAb-UL50, mAb-emerin, and mAb-β-actin. Ig-HC, cross-reactive band for immunoglobulin heavy chain.