Fig. 6. Molecular characterization of the megalin/lrp2 knock-down. Wild-type embryos were injected with buffer (WT) or with two different megalin/lrp2 splice-variant MOs (megMO1 or megMO2). Embryo extracts were generated at 48 hpf and analyzed by (A) RT-PCR or (B) western blotting of membrane fractions. megMO1 and megMO2 resulted in aberrant splicing of the megalin/lrp2 transcript as evidenced by truncated PCR products of the (A) cDNA encoding the transmembrane and intracellular portions of the receptor and (B) the absence of megalin/LRP2 immunoreactivity compared to controls. (C) Morphology of wild-type, megalin/lrp2 and dab2 morphants at 72 hpf. No gross abnormalities of the different morphants were apparent.