Figure 2

Expression of RPS10/rNef complexes in primary PBMCs, PBLs and MDMs treated with rNef.

(A) TotalTotal cellular extracts from PBMCs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb or an anti-Nef mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb or an anti-RPS10 mAb. Results are representative of three independent experiments. (B) Cytoplasmic and nuclear extracts from several cell lines (Vero cells, U937 cells), PBLs and MDMs treated with rNef (100 ng/ml) for 2 hours were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. The controls are input controls from total cell lysates (for IP RPS10 and mouse IgG control). Results are representative of three independent experiments. (C) Cytoplasmic and nuclear extracts from U937 cells treated with increasing concentrations of rNef (100–1500 ng/ml) for 2 hours or mock-treated were immunoprecipitated with an anti-RPS10 mAb. Immunoprecipitated material was analyzed by western blotting with an anti-Nef mAb. Results are representative of three independent experiments. β-actin and TBP (TATA binding protein) detection represents input loading controls of the lysates which were used in binding reactions.