Fig. 6. Lysosomal distribution and function during the formation and/or clearance of the FAG. (A-C) Vero cells were treated with Jpk (+Jpk, 50 nM) for different time periods, as indicated in the panels. Cells were double-stained with TRITC-phalloidin (red) and anti-LAMP2 antibodies (green). Lysosomes in cells containing a FAG show either a uniform distribution or accumulate around the FAG (arrow in B). At 48 hours after Jpk treatment, an increase in lysosomal staining was observed despite the absence of a FAG. (D-F) Vero cells were treated with Jpk (+Jpk, 50 nM for 6 hours) and then co-incubated with bafilomycin A1 (Baf, 100 mM; +Jpk +Baf; D,E) or pepstatin A (Pep, 10 µM; +Jpk +Pep; F). Notice that the dysfunction of the lysosomal activity prolonged the lifespan of the FAG. (E) Enlargement of the boxed area in D, in which the co-staining with TRITC-phalloidin and anti-LAMP2 antibodies revealed that Baf treatment traps lysosomes in an F-actin net (asterisks) that is structurally different from the FAG (arrowhead). Scale bars: 10 µm.