Perhexiline maleate

The peroxisome PPAR-α agonist bezafibrate (Cayman Chemical, MI, USA), the PPAR-α antagonist GW6471 (Cayman Chemical, MI, USA), and/or the CPT1 inhibitor perhexiline maleate (Cayman Chemical, MI, USA) were cultured with Jurkat cell in vitro at the indicated concentrations^{41 (link), 42 (link)}. Catalase (Sigma-Aldrich) was used to inhibit ROS production at 1000 U/mL^{8 (link)}. perhexiline maleate was injected intraperitoneal (i.p.) to inducible liver-specific MYC oncogene transgenic mice (MYC-ON) at 8 mg/kg in 100 µL 50/50 solution of PBS/DMSO three times per week for a total of 5 weeks^{27 (link)}.

C57BL/6 mice were purchased from Charles River Laboratories (VA, USA) at 8–10 weeks of age. NAFLD was induced by feeding mice the MCD diet (catalog number 960439, MP Biomedicals), or custom made high C18:2 (12%) or low C18:2 (2%) diet (Research Diets) as previously described^{8 (link)}. Liver-specific inducible MYC oncogene transgenic mice (MYC-ON) have been previously used by our group where MYC expression in the liver was activated by removing doxycycline treatment from the drinking water of 4-week-old double transgenic mice for TRE-MYC and LAP-tTA^{8 (link), 38 (link), 39 (link)}. The CPT inhibitor, perhexiline maleate (Cayman Chemical, MI, USA) was given intraperitoneally to MYC-ON mice at 8 mg/kg in 100 µL 50/50 solution of PBS/DMSO three times per week from week 6 to week 11, during which time the mice were fed with the MCD diet^{27 (link)}. MYC-ON mice were injected i.p. with 100 µL of vehicle without perhexiline as control. Mice were approximately 20 g at time of injections. Upon removal of the liver, sections of the liver were fixed with 10% formaldehyde and sent to Histoserv (Germantown, MD, USA) for hematoxylin and eosin (H&E) staining. All animal experiments were performed according to the institutional guidelines and approved by a NCI-Bethesda (Bethesda, MD, USA) Institutional Animal Care and Use protocol.