Figure 8.

Survivin overexpression rescued PKD phenotypes in zebrafish. (a) Zebrafish embryos were scored for phenotypic observations at different developmental stages. Shown here are representative images of zebrafish at 24, 48 and 72 hours-post-fertilization (hpf) injected with either control morpholino (MO), pkd2 MO, pkd2 MO plus survivin mRNA, survivin mRNA alone, or pkd2 MO plus VEGF. Abnormal phenotypes associated with pkd2 MO injections such as curly tail and renal cyst were rescued by survivin mRNA or VEGF injection into zebrafish embryos. Representative images of 48-hpf zebrafish sections are shown. The sections were stained with H&E (bottom panels), and arrows point to pronephric structures. (b) RT-PCR was performed to examine zebrafish (zf) and human (hu) transcript levels for survivin and to confirm pkd2 knockdown. Human survivin was introduced through mRNA injection. α-tubulin was used as a loading control. (c) Expression levels of survivin were analyzed in all the groups, in which zebrafish and human survivin can be recognized by the same antibody. (d) Analyses of individual chromosomes were performed in all groups of fish embryos to study chromosomal number. Black bar=500 μm; red bar=50 μm. All quantification and statistical analyses on Western blot, RT-PCR and polyploidy level are presented in the Supplementary Materials.