Aav8 cmv pi egfp wpre bgh

All animal protocols in this study were approved by the University of Pennsylvania’s Institutional Animal Care and Use Committee (IACUC) and Princeton University’s IACUC. For diet studies, 4-week-old male mice were placed on either a regular chow diet (Lab Diet 5010) or a high-fructose chow diet (Teklad TD.89247) for indicated lengths of time. Weights of mice kept on each diet were taken weekly. For drinking water studies, mice were provided with regular tap water (filtered through a 0.22 μm filter) or a 15% (w/v) fructose:15% (w/v) glucose (Sigma F3510, G8270) in tap water (filtered through a 0.22 μm filter). To deplete the gut microbiome, mice were given a daily 10 μL/g body weight oral gavage consisting of 1 mg/mL ampicillin, 1 mg/mL gentamicin, 0.5 mg/mL vancomycin, 1 mg/mL neomycin, 1 mg/ml metronidazole in a 0.9% NaCl solution for 7–10 days. Studies were controlled to mice given the same 0.9% NaCl solution without antibiotics. To knockdown Acss2, 6–8 week-old male mice were injected via tail vein with 2.0 × 10¹¹ GC/mouse AAV8.U6.shAcss2.CMV.eGFP.SV40 (University of Pennsylvania Vector Core) or AAV8.CMV.PI.eGFP.WPRE.bGH (Addgene) as control; experiments were performed 1 week after injection.