Fig. 6

Rescue of the pronephric uptake defect in OCRL1 deficient embryos.

A. Lateral view of a wild-type zebrafish embryo co-injected with cmlc2:GFP and enpep:OCRL1a. B. Schematic diagram of OCRL1 showing domain organization and localization of deletion and point mutants used in rescue experiments. C. RT-PCR detection of OCRL1 mRNA in WT, ocrl^-/- embryos or ocrl^-/- embryos expressing the indicated OCRL1a constructs. eIF1α was used as a control. The vector only lane is a positive control for the OCRL1 PCR, and lacks eIF1α. D. Images of pronephric uptake of Alexa 488-10 kDa dextran (green) in wild type (WT), ocrl^-/- embryos or ocrl^-/- embryos expressing OCRL1a, catalytically inactive GFP-OCRL1a (D480A), OCRL1a unable to bind clathrin (ΔLIDID ΔLIDLG), Rab GTPases (F727V), F&H domain proteins such as APPL1 and IPIP27A/B (W798A), or OCRL1a harbouring the Lowe syndrome mutation L746P or Dent-2 mutation P858L. The pronephric tubules are indicated with a dashed line. E. Quantification of pronephric uptake of Alexa 488-10 kDa dextran in each of the indicated embryo types. Data are presented as the mean ± SEM. Statistical analysis was performed using the Pearson’s chi-squared test. ***p < 0.0001, **p < 0.001, *p < 0.01.