Fig 1. Phenotype and nephrin expression in nephrin depleted zebrafish.

(A) Three categories of phenotypes were defined in nephrin morpholino injected embryos: embryos without edema (normal), embryos with visible pericardial edema (white arrow) and dysmorphic or dead embryos. Pictures were taken at 5 dpf. (B) 80 embryos per condition were live-screened at 5 dpf and assigned to a phenotype category. Increasing concentrations of the injected nephrin morpholino were associated with an increasing percentage of embryos with pericardial edema, but also with increased numbers of severely dysmorphic or dead embryos. (C) RT-PCR was performed using total RNA extracted from nephrin morpholino (100 μM) and control morpholino injected embryos at 24, 48 and 72 hpf. Reduced expression of normal nephrin (389 bp) was found in the nephrin depleted versus control embryos. Moreover, injection of nephrin morpholino induced alternative splicing resulted in 272 and 1070 bp fragments, resulting from an exon deletion and a retained intron, respectively. (D) Quantitation of nphs1 RNA expression in control versus nephrin morpholino injected larvae after 24, 48 and 72 h of injection. M, marker; Mo, nephrin morpholino injected; Co, control morpholino injected.