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The NO response was quantified by measuring urinary NO x (sum of nitrate and nitrite) excretion by using a colorimetric enzymatic kit (Roche Diagnostics), as described elsewhere[55]

Phenotypic analysis of F0 crispants and stable mutant embryos After anesthesia using 0.02% tricaine, thyroid development was monitored in live zebrafish at 55 hpf, 80 hpf, and 6 dpf by visual inspection using a M165 FC fluorescence stereomicroscope (Leica)