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. 2010 Jun 23;84(17):8596–8606. doi: 10.1128/JVI.00298-10

FIG. 1.

FIG. 1.

UL20p interacts with gB in virus-infected cells. Vero cell monolayers were infected with either the V3032 virus (having only gK tagged with the ProtC epitope) or the V3024 virus (having UL20p tagged with the 3×FLAG epitope and gK tagged with the ProtC epitope) or were mock infected. Cellular extracts were prepared at 24 hpi and in the presence of gB or UL20p and were visualized with Western immunoblots before or after immunoprecipitation with either anti-gB (α-gB) or anti-UL20p antibody. (A) Immunoblots probed with the anti-UL20p antibody (anti-3×FLAG). Lane 1, antigen from mock-infected cells; lane 2, V3032-infected cell extracts; lane 3, V3024-infected cell extracts; lane 5, mock-infected cell extracts immunoprecipitated with gB (gB beads) and then reacted with anti-UL20p antibody; lane 6, V3032-infected cell extracts immunoprecipitated with gB and reacted with anti-UL20p antibody; lane 7, V3024-infected cell extracts immunoprecipitated with anti-gB and reacted with the anti-UL20p antibody. Lanes 9, 10, and 11 are the same as lanes 5, 6, and 7, respectively, except that the initial immunoprecipitation was performed with anti-UL20p antibody and immunoblots were probed with anti-UL20p antibody. Lanes 4, 8, and 12 demarcate the relative positions of molecular mass markers (10 to 250 kDa; Bio-Rad). (B) Exact same arrangement of reactivity patterns shown above (A) except that the immunoblot was probed with anti-gB antibody.(C) Immunoblot probed with anti-gD antibody. Lanes 9, 10, and 11 are lysates from mock-infected cells, V3032-infected cells, and V3024-infected cells probed with the anti-gD antibody, respectively. Lanes 1, 2, and 3 are the same samples used for lanes 9, 10, and 11, respectively, except that the samples were first immunoprecipitated with anti-gB antibody prior to probing with the anti-gD antibody. Similarly, samples in lanes 5, 6, and 7 are the same samples shown in lanes 9, 10, and 11, respectively, except that the samples were first immunoprecipitated with anti-FLAG antibody and subsequently probed with anti-gD antibody.