Fig. 1.

Immunoprecipitation of mRFP-gB and VP26-Venus from YK614-infected cells. Vero cells were infected with either wild-type HSV-1 strain F, YK614 (carrying the mRFP-gB and VP26-Venus sequences), F-BAC gB−, which does not express gB (7), or F-US9−/GFP, which expresses GFP (21, 22). After 6 h, the cells were radiolabeled with [³⁵S]methionine-cysteine for 150 min; then the cells were lysed in NP-40–DOC buffer containing 2 mg/ml BSA and 1 mM PMSF. Extracts were centrifuged at high speed to remove insoluble debris and were then mixed with anti-gB monoclonal antibody (MAb) 15βB2 (A) or with rabbit anti-GFP antibodies (B). Then protein A-Sepharose was added, and gB or GFP was immunoprecipitated as described previously (27). The positions of gB, mRFP-gB, GFP, and VP26-GFP, as well as those of molecular mass markers of 220, 97, 66, 46, and 30 kDa, are indicated.