Fig. 2. Chronological acquisition of CI-MPR, CD63 and Hck during the maturation of phagosomes containing zymosan. MDMs were infected with zymosan under synchronized conditions. At different post-infection times (5 (a,b), 10 (c,d) or 30 (e,f) minutes), cells were fixed and permeabilized in methanol. Double immunostaining was performed using Abs directed against CD63 and CI-MPR or Hck. We focused on phagosomes, and the presence of the markers at the phagosomal membrane was determined by a fluorescent ring (see arrows). (A) CI-MPR, a marker of late endosomes (a,c,e) was delivered to phagosomes before CD63 (b,d,f) (arrows). (B) CD63 (a,c,e) was delivered to phagosomes before Hck (b,d,f) (arrows). Occasionally Hck and CD63 were found on the same phagosome (arrowhead). Data are from one experiment that is representative of two to four independent experiments. Bar, 7 µm.
