Figure 5.

Co-IP–based interaction analysis of domain swap constructs between HCMV pUL53 and EBV BFLF2. A, schematic representation of the domain swap constructs generated between pUL53 (blue) and BFLF2 (green); numbers represent the amino acid positions. B, summarized data of the co-IP analysis with domain swap pUL53::BFLF2 proteins in their interaction with HCMV pUL50 or EBV BFRF1, respectively. C-F, 293T cells were transiently transfected with the expression plasmids coding for domain swap proteins as: C and E, FLAG-tagged constructs containing the main globular domains; D and F, GFP fusions replacing the globular domains. At three days post-transfection, cells were lysed and HA-tagged proteins were immunoprecipitated using anti-HA antibodies as indicated. C and D, test for interaction with pUL50-HA. E and F, test for interaction with HA-BFRF1. Co-IP procedures were performed as described previously (32). In each panel, at least three independent experiments have been performed; one representative set of data are shown.