Figure 3.

Kinetics of acquisition of early endosomal EEA1 or late endosomal LAMP-1 by phagosomes containing either Afipia or zymosan. APCs (A) and zymosan-containing phagosomes (B) (please note different scales in A and B). Experiments were done as in Fig. 2 with a spin at 160 × g for 30 min in a tabletop ELISA plate centrifuge with swingout rotor at 4°C, followed by removal of media and addition of fresh 37°C media (being set as 0 min), plus chase for up to 120 min. Glass slide preparations were fixed at the indicated times and rings of antibody staining by using anti-LAMP-1 (□) or anti-EEA1 (●) were counted positive in confocal microscopy. For every time tested, added colocalization frequencies are indicated as well (○), equaling the number of phagosomes that are part of the endocytic system. Results are from three independent experiments with a minimum of 50 phagosomes counted per time and sample type.