Journal: PLoS Pathogens
Article Title: Binding of HIV-1 gp120 to DC-SIGN Promotes ASK-1-Dependent Activation-Induced Apoptosis of Human Dendritic Cells
Figure Lengend Snippet: Activation of gp120-primed DC reduced the expression of Bcl2 and activated Akt, and the induction of cell apoptosis is ASK1-dependent. ( A ) moDC treated with immune-complex gp120 ADA were exposed to CL40L Tf or mock Tf, or LPS, TNFα, IL-1β for 3 d, and cellular proteins of moDC (recovered from coculture) were extracted for Western blotting analysis. MoDC treated by anti-His Ab (used to cross-link recombinant gp120 ADA ) were used as a control (control DC). *p-ASK1 represent results from untreated moDC (lane 1) and DC treated as indicated but no gp120 pulsing (lanes 2–6), whereas #p-ASK-1 represents results from pre-treatment of anti-DC-SIGN mAbs. Data are representative of 3 experiments. ( B ) moDC were transfected with siRNA against human ASK1 or control siRNA (scrambled) before pulse with immune-complex gp120 and subsequent exposure to CD40L Tf, LPS, TNFα or IL-1β. Data are expressed as mean ± SD from 3 experiments. **p
Article Snippet: Figure S2 EndoH treatment of recombinant gp120 abolished its binding to moDC. (A ) Recombinant gp120ADA supernatant was treated overnight with 25 KU of EndoH/ml as described (Hong PWP et al, J Virol 2002;76:12855–12865), and the treated and the untreated gp120ADA supernatant were subjected to Western blot assay by polyclonal rabbit anti-gp120 Ab (Sino Biological Inc).
Techniques: Activation Assay, Expressing, Western Blot, Recombinant, Transfection