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Fig. 1. Agrp overexpression promotes obesity in casper zebrafish. (A) Schematic of generation of zebrafish agrp cDNA (zAgRP1)-overexpressing casper zebrafish (mitfa:BRAFV600E, p53−/−, mitfa−/−, mpv17−/−). Created with Biorender.com. WT, wild type. (B) Weight of both male and female F0 fish combined over 6 months. (C,D) Weights of male (C) and female (D) F0 mosaic fish at 120 days post-fertilization (dpf). Fish were separated into equal numbers at 1 month, and their weights were measured at the indicated time points. n=5-10 fish per genotype per biological replicate. Data are the averages of three biological replicates. (E,F) Weights of male (E) and female (F) fish of the F3 stable line at 120 dpf. Data are the weights of all the fish from two clutches from separate single male and female pairs of fish across six separate tanks. n=35-60 fish per group. zAgRP1 fish and wild-type siblings were housed in the same tanks and identified via GFP fluorescence. (G) Representative images of male and female zAgRP1 fish and wild-type siblings. (H) Histology of abdomen from 7-month-old F3 male and female zAgRP1 or wild-type siblings. Representative images are shown. Fish were fixed in PFA, sectioned and stained with H&E. (I,J) Quantification of adipose tissue (AT) per section from male (I) and female (J) fish. Three fish per sex per genotype and two sections per fish were analyzed. (K,L) Quantification of individual adipocyte size of male (K) and female (L) fish. (M) Histology of liver from 7-month-old male F3 male and female fish. Fish were fixed in PFA, sectioned and stained with H&E. Three fish per genotype per sex were sectioned for histology. *P≤0.05, **P≤0.01. Welch's t-test. Scale bars: 5 mm (G), 1 mm (H, top row), 500 μm (H, bottom row), 50 μm (M).