Various molecular mechanisms of unconventional secretion of fibroblast growth factor 2 and galectin-1 have been proposed. A non-vesicular pathway that is based on direct translocation across the plasma membrane has been described. In other studies, however, release into the extracellular space of cell-derived vesicles was implicated in both FGF-2 and Gal-1 secretion. Such vesicles were proposed to originate either from plasma membrane shedding or by the release of exosomes. Employing an inhibitor of plasma membrane blebbing and based on a quantitative biochemical analysis of cell culture supernatants for vesicles potentially carrying FGF-2 or Gal-1, we demonstrate that both FGF-2 and Gal-1 are not exported by shedding of plasma membrane-derived vesicles.