FIG. 5.

Surface localization of p10 constructs. Live cells transfected with various p10 constructs were treated with anti-HA monoclonal antibodies to detect the presence of the surface-localized N-terminal HA epitope tags on the p10 constructs. Surface-bound anti-HA antibodies were detected by immunofluorescence with fluorescein isothiocyanate-conjugated goat anti-mouse IgG. A subset of the p10 constructs are shown. All of the p10 constructs, assessed in the same manner, yielded similar results. Images were captured under identical parameters. Magnification, ×630; bars, 20 μm. The specificity of the surface-staining protocol was confirmed by using a p10 construct with HA tags in the C-terminal endodomain, as previously described (45), and is evident from the low level of background staining of the surrounding nontransfected cells in the confluent monolayer.