Fig. S2

PKD1 expression silencing by two splice-blocking morpholinos and PKD1 mRNA injection in zebrafish embryos. A–A2, Expression silencing of PKD1 in 24 hpf zebrafish embryos using two splice-blocking morpholinos (SB-Mo I5 and I6). RT-PCR analysis of 2 ng control morpholino (Co-Mo) and 500 pg SB-Mo I5 (A) or 500 pg SB-Mo I6 (A2) injected embryos. The upper signal represents the wild-type (wt), the lower signal the morphant (mo) product. Injection of SB-Mo I5 or I6 generated a substantial loss of the wild-type splice product indicating the functionality of both PKD1 morpholinos. B–B2, PKD1 antibody stainings of cross-sections of 48 hpf embryos injected with CoMo (2 ng) (B) or SB-Mo I5 (500 pg) (B2) indicated reduced PKD1 expression (red colour) in PKD1 morphant embryos. Exposure time was 840 ms for all images. C, Injection of 100 pg human PKD1 sense RNA into zebrafish embryos led to an enhanced expression of PKD1 protein. C2, Quantification of (C) by densitometry, showing three independent experiments with S.D. D, Western blot analysis of 24 hpf embryos after injection of SB-Mo I5 (500 pg) or SB-Mo I6 (500 pg) indicated strong reduction in PKD1 protein expression as compared to Co-Mo - injected (2 ng) embryos. D2, Quantification of (D) by densitometry, results represent means of three independent experiments with S.D. E, Expression of PKD1 in tg(fli1:EGFP) zebrafish endothelial cells at 24 hpf as shown by RT-PCR of EGFP purified endothelial cells. Scale bars: 25 μm.