Immunoblot analysis of cell signaling induced by C2-2b-2b. (A) Daudi cells were untreated (lane 1) or treated for 2 hours at 20nM with C2-2b-2b (lane 2), hL243γ4p (lane 3), or 734-2b-2b (lane 4) and evaluated with loading 20 μg of total protein/lane. (B) CAG cells were treated for 1 hour with C2-2b-2b (left) or 20-2b-2b (middle). The picomolar concentration of mAb-IFNα is indicated at the bottom of each lane. Phospho-Stat-1 and phospho-Stat-3 were measured with loading 15 μg of total protein/lane. (C) Daudi cells were treated for 1 hour with 0.1nM of C2-2b-2b (left) or 734-2b-2b (right), washed, and then incubated an additional 48 hours. Phospho-Stat-1 and phospho-Stat-3 were measured with loading 10 μg of total protein/lane. Time points indicate hours after washing; and U, untreated. β-actin was used to verify equal loading. Specific Ab probes are indicated to the right of each panel.