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Figure 7


Fig. 7. Tubular-clearance defects in megalin/lrp2 morphants and quantification thereof. (A,E,I) Wild-type embryos, (B,C,F,G,K,L) megalin/lrp2 morphants, (D,H,M) dab2 morphants at 72 hpf were injected with 70kDa-FD (green). Renal clearance of tracer into the pronephric duct tubular cells was evaluated after fixation and immunostaining against megalin/LRP2 (red) of whole mounts by confocal fluorescence microscopy. Renal clearance of tracer occurs in (A,I) wild-type but not in (B,C,K,L) megalin/LRP2- and (D,M) Dab2-deficient embryos. Expression of megalin/LRP2 is unaffected in dab2 morphants (H). (N) Wild-type, megalin/lrp2 morphant, dab2 morphant and dab2 rescued embryos at 72 hpf were injected with the indicated fluorescent tracers and the number of embryos exhibiting tubular accumulation of tracers evaluated by fluorescence microscopy. Data of all animals injected are given in percent. 70kDa-FD: wild type, 45/55 embryos (81.8%); megMO1, 2/24 embryos (8.3%); megMO2, 3/29 embryos (10.3%), dab2MO, 3/29 embryos (10.3%); dab2 rescue, 26/48 embryos (54.2%). 10kDa-RD: wild type, 46/52 embryos (88.5%); megMO1, 4/27 embryos (14.8%); megMO2, 5/29 embryos (17.2%); dab2MO, 5/39 embryos (12.8%). Cy2-RAP: wild type 42/46 embryos (91.3%); megMO1, 3/25 embryos (12.0%); megMO2, 3/22 embryos (13.6%); dab2MO, 3/31 embryos (9.7%). Bar, 100 µm.





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