Fig. 5

Fig. 5 The hepatic metabolic defects in sec31anju221 adult fish can be alleviated by thyroid hormone treatment. (A) Left: whole-mount RNA in situ hybridization for thyroglobulin in the thyroid gland of adult fish. Right: representative immunofluorescence of thyroxine (T4, purple) in the thyroid follicle lumen of wild-type and sec31anju221 fish (n=5 zebrafish per group). Thyrocyte identity is confirmed by DAPI counterstaining (white). Scale bars: 500 ?m (left); 50 ?m (right). (B) Serum thyroxine (T4) concentrations in wild-type and sec31anju221 fish (n=5). Data are mean±s.e.m. (C) Western blot analysis and relative quantification of Dio1 from adult wild-type and sec31anju221 fish liver (n=3). (D) Western blot analysis and relative quantification of Dio1 from adult wild-type, vehicle-treated sec31anju221 and T4-treated sec31anju221 fish liver (n=3). (E) T4, triglyceride and glycogen levels in fish liver were measured with enzymatic colorimetric assays. n=9, 9 and 8 for control, vehicle-treated sec31anju221 and T4-treated sec31anju221 fish, respectively. Each data point represents an individual animal, with bars representing mean±s.e.m. Comparisons between groups in B and C were performed using two-tailed unpaired Student's t-tests, and comparisons between groups in D and E were performed using one-way ANOVA with Tukey's multiple comparisons test. ns, not significant; *P<0.05; **P<0.01; ****P<0.0001. (F) Representative photographs of H&E, ORO and PAS staining of liver biopsy samples. Arrowheads indicate oil droplets. Images are representative of five fish per group. Scale bars: 50 ?m.