Fig. S2

Role of MC4R, AgRP and POMC in Zebrafish Growth.

(A-C) mc4r-/- fish exhibit increased growth by 42 dpf. Fish from a sa0149 mc4r +/- intercrossing were allowed to grow to adulthood. Body lengths (jaw to trunk terminus, tail fin excluded) of juveniles were measured at 42 days post fertilization (A). Female (B) and male (C) adult zebrafish were also measured at 8 months. Bars indicate mean±s.e.m. Results were analyzed by one way ANOVA followed by Tukey post test. (N=5-53, *P<0.05;***P<0.001). (D) Partial rescue of agrp MO inhibition of somatic growth by co-injection of pomca MO. Body lengths at 5 dpf following injection of wild type zygotes with 2 ng MO indicated, plus 0.4ng pomca ATG antisense MO showing partial rescue of growth. Bars indicates mean±s.e.m. n= 42, 42, 39. Statistical significance tested by one way ANOVA followed by Tukey post test. (***, p<0.001). (E-F) Developmental expression of melanocortin receptor and ligand genes. Relative expression levels of (E) mc1r, mc3r, mc4r, mc5ra, and mc5rb, and (F) agrp and pomca were analyzed by QPCR. ~200 WT zebrafish zygotes were raised and sacrificed at days indicated for RNA extraction and cDNA synthesis. 4dpf group was normalized to 100% and all gene expression was normalized to the house-keeping gene, ef1a or β-actin. Results are expressed as mean + s.e.m.