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Fig. 3
Depletion or knockout of Pacsin2 in zebrafish larvae impairs renal tubule endocytosis. (A) Schematic representation of dextran reabsorption assay performed on zebrafish larvae. (B) Top, representative image of 4?dpf pacsin2+/? (left) and pacsin2?/? (right) larvae 1.5?h post-injection with 10?kDa dextran-A488, displaying normal and no uptake phenotypes, respectively. Scale bar: 25?µm. The blue dashed line indicates the position of the proximal tubule. Bottom, the percentage of larvae showing normal, low or no uptake phenotypes in pacsin 2+/? and pacsin 2?/? larvae. Data were analysed using a Chi-squared test. **P<0.01. n=67 (pacsin2+/?) and 41 (pacsin2?/?). (C) Top, representative images of 3?dpf control larvae showing normal accumulation of 10?kDa dextran-A488 at 1.5?h post-injection and morphant embryos displaying low or no uptake phenotypes, respectively. The blue dashed lines outline the position of the proximal tubule. Scale bar: 50?µm. Bottom, quantification of uptake phenotypes of control, E2MO and E4MO morphant larvae. Data were analysed using a Chi-squared test. **P<0.01, ***P<0.001, ****P<0.0001. n=28-84 larvae per genotype. (D) Top, representative images of cmlc:GFP (top row, mock) and enpep:pacsin2 (bottom row) rescued pacsin2+/? and pacsin2?/? larvae. The blue dashed lines outline the proximal tubule. Scale bar: 60?µm. Bottom, percentage of total larvae scored for uptake phenotypes. Data were analysed using a Chi-squared test. **P<0.01. n=23-56 larvae per genotype.