Fig. 1

The clathrin complex is required for slit diaphragm protein endocytosis. Immunofluorescence labeling for Pyd (red), with DAPI (blue) nuclear stain of adult fly nephrocytes, visualized by confocal microscopy. Scale bar: 1 μm. a–a″ Wild type (Control) adult fly nephrocytes exhibited Pyd (red) localization by immunofluorescence labeling that was tightly localized to the cell margin, defining a highly regular, continuous circumferential ring when examined by confocal microscopy (a, medial optical section). Visualization of Pyd at the cell surface plane showed a highly regular and uniform pattern of distribution (a′–a″). Cell nuclei were labeled by DAPI (blue). Scale bar: 1 μm. b Silencing of Arf79f (b–b″) did not cause intracellular accumulation of Pyd proteins. But on the cell surface section, the distance between the lines of Pyd fingerprint become wider. c–d Silencing of Chc (c–c″) or Clc (d–d″) genes of the clathrin complex resulted in a severe disruption of normal Pyd localization at the nephrocyte cell margin and the cell surface. Intracellular Pyd showed a distribution with aggregate-like concentrates. The fingerprint pattern of Pyd at the cell surface was also disrupted: the highly organized and evenly spaced lines were gone, replaced by irregular short lines or aggregates