Fig. 3.

Apilimod treatment modestly increases the level of well-known exosome-associated proteins in exosomal fractions. a PC-3 cells were seeded on 10 cm plates and pretreated with apilimod (0.5 µM) or DMSO (0.1 %, as control) for 2 h before collection of exosomes for 18–19 h in the presence of inhibitors. Exosomes were isolated by ultracentrifugation and lysed in total 80 µl (lysis buffer plus sample buffer), before 25 µl of this mixture was loaded per well and run on SDS-PAGE gel. Each treatment was run in duplicates per experiment. Representative Western blot and quantification of proteins from exosomal fractions from several independent experiments. b Representative Western blot and quantification of cellular proteins from several independent experiments. The results are plotted as dot plots showing the value obtained for each individual experiment as a single dot + mean values + standard error of the mean. *; P < 0.05 versus control, #; P < 0.05 versus apilimod, n = 3–4