7 12 dimethylbenz a anthracene dmba

Male wild-type C57BL/6J mice (#000664, 9–10 weeks/old), Cybb^fl/fl mice (#031777), and Rosa^NICD mice (#008159) were from Jackson Laboratory (Bar Harbor, ME) and allowed to adapt in the animal facility for 1 week prior to random assignment to experimental cohorts. Wwtr1^fl/fl mice^{20 (link)}, backcrossed to C57BL/6J, were provided by Dr. Eric Olson (University of Texas Southwestern). The mice were fed a diet containing sugar water (23.1 g fructose/L and 18.9 g glucose/L), palmitate, and 1.25% cholesterol (“NASH diet”; Teklad, TD.160785 PWD), which induces NASH after 16 weeks^{8 (link)}. All AAV8-viruses were injected by tail vein (2×10¹¹ genome copies/mouse) as indicated in the figure legends. For the DMBA model, 50 μl of 0.5% DMBA (7,12-dimethylbenz [a]anthracene, Sigma) in acetone was administered to the dorsal surface on postnatal day 4–5^{21 (link)}; the NASH diet was begun at weaning (3 wks/o). Animals were housed in standard cages at 22°C in a 12–12-hour light-dark cycle in a barrier facility. For mouse HCC, the predetermined endpoint was tumor weight estimated to be <10% of body weight. All animal experiments were performed in accordance with institutional guidelines and regulations and approved by the Institutional Animal Care and Use Committee at Columbia University.