Flow cytometry analysis of Env-expressing cells labeled with Halo ligands or anti-Env monoclonal antibody.

<p>(A–C) Flow cytometry analysis of Env expression level using different staining strategies. 293FT cells were transfected with expression vectors for tethered Env (HXB2-TM11D-Halo, red line; JRFL-TM11D-Halo, blue line), untethered Env (HXB2-WT, red dashed line; JRFL-WT, blue dashed line), or Mock DNA (grey shade). Cells were stained with membrane-impermeable HaloTag AF488 ligand (A), membrane-permeable HaloTag Oregon Green ligand (C) and anti-Env V3 antibody V3-G2-25 (B). Histograms are representative results from three independent experiments. (D) Positive staining rate of HaloTag labeling and anti-Env antibody immunolabeling of cells transfected with HXB2-TM11D-Halo (solid red bar), HXB2-WT (red shade bar), JRFL-TM11D-Halo (solid blue bar), JRFL-WT (blue shade bar) or Mock (solid gray bar). Error bars represent standard deviations of the results from three independent experiments.