Fab-based inhibitors reveal ubiquitin independent functions for HIV Vif neutralization of APOBEC3 restriction factors
Immunoblot analysis of cellular A3 levels and packaging into VLPs in presence of Vif and Fab derived scFvs. Env-deficient NL4-3 HIV (either Vif+ or Vif-) was co-transfected into HEK293T cells with HA-tagged A3 and a gradient of Flag-tagged scFv3C9 (A-C) or scFv1D1 (D-F). The A3 proteins, Fabs, and Vif were visualized by WB using anti-HA, anti-Flag, and anti-Vif. p24 and GAPDH were used as loading controls for the VLP and cell fractions, respectively.