Fig. 6.

VRC01 effects on HIV envelope glycoprotein functional trimer. (A) Western blot analysis of Env from supernatants of 293T cells transfected with cleavage competent gp160 JRFL or JRCSF Env plasmid DNA. HIV-1 gp120 shedding upon binding of ligands, including sCD4, b12, and VRC01, from the Env trimer on the surface of 293T cells was evaluated by probing the blot with the anti-gp120 human monoclonal antibody 39F (V3 specific). (B) Effect of VRC01, b12, and sCD4 on gp41 MPER-directed antibodies 2F5 and 4E10 binding to trimeric Env. 293T cells transfected with JRFL-ΔCT Env expression plasmid were stained with biotin-labeled 2F5 or 4E10. Binding of 2F5 or 4E10 to JRFL Env in the presence of the CD4bs ligands VRC01, b12, and sCD4 was detected with streptavidin-APC conjugate. MFI, mean fluorescence intensity.