Time course analysis of NS3 intracellular trafficking. (A-B) CLSM examinations (central optical sections) of DCs that were allowed to take up NS3 for 15 minutes and then were washed and chased with medium for the indicated time intervals. After staining with anti-NS3 Ab, cells were analyzed for the expression of selected different markers to evaluate the localization of internalized NS3 Ag (red) into early endosomes (EEA-1 and Rab5), late endosomes (Rab7), recycling vesicles (Rab11), and ER (calnexin), all detected in green. Cells were counterstained with anti–NOX-2 and MHC-I Abs to evaluate the delivery of NOX-2 (green) and MHC-I (green) molecules to NS3⁺ organelles. Colocalization areas were detected in yellow. Data are representative of 7 independent experiments.