Packaging of HSV1 pUL36 proteins into virions. (A) Vero cells were infected with HSV1(17+) at an MOI of 10 PFU/cell or mock treated, harvested at the indicated time points, and separated on a linear 7.5 to 18% SDS-polyacrylamide gel. The proteins were transferred to a nitrocellulose membrane and probed with antibodies against pUL36 aa 1408 to 2112 (middle, pAb 146), VP5 (pAb NC-1), or actin (MAb 1501). (B) Vero cells infected with HSV1(17+) at an MOI of 10 PFU/cell for 12 h (cells) or gradient purified HSV1(17+) secreted from infected BHK cells and purified on a linear Nycodenz gradient (virus) were lysed; and the proteins were separated on linear 5 to 15% SDS-polyacrylamide gels, transferred to nitrocellulose membranes, and probed with antibodies directed against pUL36 aa 33 to 44 (Nterm, pAb R28), pUL36 aa 1408 to 2112 (middle, pAb 146), pUL36 aa 3048 to 3057 (Cterm, pAb R29), VP16 (MAb LP1), or VP5 (MAb H1.4).