Fig. 6

Intracellular trafficking of endocytosed AFP. a nAFP and tAFP were Alexa Fluor 488-labeled. Human monocyte-derived DC were co-cultured with AFP (10 μg/ml) for 1 hr at 4 °C or 37 °C, and analyzed by flow cytometry. b DC were co-cultured with fluorescently-labeled nAFP for 2 hr, then chased with media alone for the indicated times. Cells were then fixed, stained for actin (described in Materials and Methods), and analyzed by confocal microscopy. c DC were incubated with fluorescently-labeled nAFP for 2 hr, then chased with media alone for the indicated times. Cells were then fixed and stained for EEA-1 (early endosomes), LAMP-1 (late endosomes/lysosomes), KDEL (endoplasmic reticulum/ER), and golgin-97 (Golgi) (all in red). All images are representative of three independent experiments performed and were taken using a 63x objective