FIG. 2.
Immunolocalization assay of cell surface-displayed His6-tagged ecotropic Env. (a) 293T cells. (b) Mock transfection of 293T cells transfected with E/A-PRR. (c) 293T cells transfected with pRVHis_e encoding the His6-tagged Env. (d) Histogram of the cells expressing the His6-tagged Env on their surface. Cells were collected 48 h posttransfection and stained with mouse anti-Penta-His antibodies as the primary antibody (1:50 dilution ratio) and goat anti-mouse IgG FITC conjugate as the second antibody (1:100 dilution ratio). After staining, the samples were applied to flow cytometry immediately. All the samples contained 106 cells. The FL2 channel measured autofluorescence emitted from the cells, which corrected the measurements from the FL1 channel (labeled as FITC channel) by adjusting the color compensation in the flow cytometry.