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Fig. S4
Phenotypic spectrum of duox and duoxa crispants. (A) Epifluorescence live imaging of Tg(tg:nlsEGFP) zebrafish larvae injected with Cas9 protein and sgRNAs targeting duox or duoxa, respectively. Ventral views of the thyroid region (anterior to the right) are shown for non-injected control fish and four crispants. Images were aquired at 55 hpf, 80 hpf, and 6 dpf. No deviation from control thyroid morphology was evident in duox and duoxa crispants at 55 and 80 hpf. By 6 dpf, hyperplastic thyroid enlargement (goiter) was detectable in many duox and duoxa crispants. (B) Whole-mount immunofluorescence staining of Tg(tg:nlsEGFP) zebrafish for EGFP and colloidal T4 at 6 dpf. Ventral views of the thyroid region (anterior to the right) are shown for non-injected control fish and different crispants displaying hypothyroidism (decreased T4 staining) that was often but not always associated with a hyperplastic thyroid enlargement. Scale bars: 50 μm.