Detection of gB-gH/gL complexes following IP from radiolabeled Ad-transduced cells.

<p>ARPE-19 cells were transduced with Ad vectors expressing HCMV glycoproteins: HCMV strain TR or strain AD169 gB, TR gH/gL or a negative control, Ad-tet-trans (tet), as indicated at the top part of each panel. At 20 hrs post-transduction, the cells were radiolabeled with ³⁵S-cysteine-methionine for 4 hrs. The cells were lysed with IP buffer containing 1% NP-40 and the proteins immunoprecipitated (IP’d) then analyzed by SDS-PAGE under reducing conditions. (A) Lysates were IP’d with anti-gH MAbs: 14-4b or AP86. (B) Lysates were IP’d with rabbit anti-peptide sera specific for gL in the absence of the gL peptide used to produce the antibodies (left panel) or with that peptide present in the IP (right panel). (C) Lysates were IP’d with the human anti-gB MAb 758. Molecular mass (MW) markers are indicated on the left.