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. 2003 Sep;77(18):9769–9779. doi: 10.1128/JVI.77.18.9769-9779.2003

FIG. 7.

FIG. 7.

Sequential addition of HA epitope tags to the C terminus of p10. The authentic p10 expression construct was modified by sequential addition of HA epitope tags to the C terminus to generate the p10-1HAC, p10-2HAC, p10-3HAC, and p10-4HAC constructs. (A) Radiolabeled cell lysates were prepared from cells transfected with each of the HA-tagged constructs, immunoprecipitated with anti-HA (α-HA) monoclonal antibody, and detected by SDS-PAGE and fluorography. The p10-1HAC construct was not precipitated by the anti-HA monoclonal antibody and was therefore precipitated with a low-avidity polyclonal anti-p10 antiserum. Asterisks indicate the electrophoretic mobilities of the various tagged p10 constructs. The relative migration of molecular mass markers (in kilodaltons) is indicated on the right. (B) Cell monolayers transfected with the same constructs were immunostained with anti-p10 or anti-HA and alkaline phosphatase-conjugated secondary antibody. Magnification, ×100. Arrows in the top two panels indicate the boundaries of syncytial foci induced by the functional 1HAC and 2HAC constructs, while arrows in the lower panels indicate single antigen-positive cells present in monolayers transfected with the nonfunctional 3HAC and 4HAC constructs.