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Fig. 5 | Cell & Bioscience

Fig. 5

From: Slit diaphragm maintenance requires dynamic clathrin-mediated endocytosis facilitated by AP-2, Lap, Aux and Hsc70-4 in nephrocytes

Fig. 5

Nephrocyte slit diaphragm structure maintenance requires clathrin. a Schematic representation of the temperature sensitive Gal80 experimental system. In brief, at 18 °C Gal80 is active, thus Gal80 inhibits Gal4 thereby preventing transcription. At 29 °C Gal80 no longer binds Gal4, thus Gal4 drives expression of Clc-IR thereby silencing expression of Clc specifically in the nephrocytes. Dorothy (Dot) promoter, nephrocyte-specific expression; tubulin (tub) promoter, ubiquitous expression pattern. be Immunofluorescence labeling for Pyd (red) of adult fly nephrocytes, visualized by confocal microscopy. Scale bar: 1 μm. bc′ At day one adult flies reared at 18 °C, wild type (Control; bb′) and Clc RNAi knockdown (Clc-IR; cc′) nephrocytes exhibited indistinguishable Pyd (red) distribution. In both, Pyd exhibited a uniform and smoothly distributed fingerprint-like pattern on the cell surface. de′ At day four, after a three-day shift to 29 °C, nephrocyte cell surface Pyd distribution in wild type (Control) flies remained unchanged, showing the characteristic fingerprint-like pattern (dd′). Nephrocytes in which the Clc gene was silenced (Clc-IR), displayed severe disruption of Pyd localization at the cell surface (ee′)

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