Zebrafish strains: zebrafish were maintained according to institutional and national ethical and animal welfare guidelines. All experiments were performed under UK Home Office licenses 40/3708 and 70/8588. The zebrafish lines Tg(-26wt1b:EGFP)li164 (link) and tmem33sh44343 (link) were used.
Zebrafish morpholino injections: Embryos were injected at the 1 cell stage. A morpholino complementary to the ATG region of zebrafish pkd2 was injected at 1 ng (5’-AGGACGAACGCGACTGGAGCTCATC-3’)42 (link). A concentration of 0.4 ng or 1 ng control morpholino (5’- CCTCTTACCTCAGTTACAATTTATA-3’) was injected in these experiments.
Image acquisition and analysis: Zebrafish embryos were imaged at 52–55hpf when cysts became visible, using a Leica M165FC stereo microscope and images were acquired using Leica LASX software. Images were quantified using FIJI v1.52i. Zebrafish renal phenotype analysis was double blinded.
Zebrafish morpholino injections: Embryos were injected at the 1 cell stage. A morpholino complementary to the ATG region of zebrafish pkd2 was injected at 1 ng (5’-AGGACGAACGCGACTGGAGCTCATC-3’)42 (link). A concentration of 0.4 ng or 1 ng control morpholino (5’- CCTCTTACCTCAGTTACAATTTATA-3’) was injected in these experiments.
Image acquisition and analysis: Zebrafish embryos were imaged at 52–55hpf when cysts became visible, using a Leica M165FC stereo microscope and images were acquired using Leica LASX software. Images were quantified using FIJI v1.52i. Zebrafish renal phenotype analysis was double blinded.
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