Fig. 5.

Obesity increases tumor growth in a sex-dependent manner in Rb1 mutant melanoma. (A) Schematic of in vivo transgenesis assay via TEAZ with the addition of an HFD. Fish were put on a control diet or HFD for 1 week. Adult casper (mitfa:BRAF^V600E, p53^−/−, mitfa^−/−, mpv17^−/−) zAgRP1 or wild-type F3 fish were injected with MiniCoopR-tdTomato, sgRB1, mitfa:Cas9 and tol2 constructs and then electroporated. Fish were analyzed for tumor initiation and area by fluorescence microscopy over 12 weeks while they remained on their respective diets. Created with Biorender.com. (B,C) Tumor growth as measured by tdTomato⁺ area over time. TdTomato⁺ area of lesions on zAgRP1 or wild-type male (B) and female (C) fish at indicated dpe. Growth curves were analyzed via mixed-effects analysis, with P=0.0344 significance when comparing the different conditions in males and P=0.2391 in females. (D,E) tdTomato⁺ area for early (21 dpe; D) and late (63 dpe; E) lesions in male zAgRP1 or wild-type fish on either a control diet or HFD. (F,G) tdTomato⁺ area for early (21 dpe; F) and late (63 dpe; G) lesions in female zAgRP1 or wild-type fish on either a control diet or HFD. (H) Representative fluorescence overlaid on brightfield images of lesions in male and female fish at the indicated time dpe. Representative images were chosen based on numerical values closest to the mean for each condition. n=14 fish per condition per sex. Data are the averages of three biological replicates. Dunnett's multiple comparisons test. Scale bars: 500 μm.