Mouse leptin neutralizing antibody

TNFrsf1a^tm1Im/TNFrsf1b^tm1Imx/J mice and wild-type C57BL/6 mice were purchased from the Jackson Laboratory. Pomc-Cre mice were produced as previously described^{47 (link)}. All mice were maintained on C57BL/6 stain background. Mice were housed in standard, pathogen-free conditions with 12 h/12 h light and darkness cycles and maintained on a normal chow diet, and male mice at adult ages (3–5 months) were used in experiments. C57BL/6 mice with diet-induced obesity were generated through 15-week high-fat diet feeding, and age-matched chowfed C57BL/6 mice were used as controls. For pharmacological injections, mice were intraperitoneally (i.p.) injected with TNF, palmitic acid, linoleic acid, cerulenin (Sigma), recombinant mouse leptin, and mouse leptin neutralizing antibody (R&D systems) at the indicated doses consecutively for 3 d. For bacterial injection, mice were intravenously injected with L. monocytogenes (ATCC) at the dose of 5 × 10⁴ CFU (colony-forming units). Mice were euthanized at indicated times post treatment, and various tissues were collected for subsequent analyses. The Institutional Animal Care and Use Committee at the Albert Einstein College of Medicine approved all the procedures.