Uptake of fluorescent nano beads into BY2-cells involves clathrin-dependent and clathrin-independent endocytosis

To follow endocytosis in BY-2 cells we made use of fluorescent nano beads. Beads with 20 nm in diameter were internalised rapidly and accumulated partially in compartments also labelled by the endocytic marker FM4-64. Studies in BY-2 cells and protoplasts revealed that larger beads (100 nm) were excluded from uptake into turgescent and plasmolysed cells while protoplasts were able to internalise beads with a diameter of up to 1000 nm. Endocytosis of beads was only partially inhibited by the clathrin-specific inhibitor Ikarugamycin and strongly blocked by wortmannin. These results imply that uptake of beads involves clathrin-dependent and clathrin-independent endocytic mechanisms and supports the hypothesis that clathrin-independent endocytosis plays a general role in plants. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.