C57bl 6j wild type control mice

Two-month-old male NLRP3 KO mice (B6.129S6-Nlrp3^tm1Bhk, which were backcrossed with C57BL6J mice for 10 generations) and C57BL/6J wild-type (WT) control mice (N = 7-8 per genotype) were obtained from Jackson Laboratories (Bar Harbor, ME, USA) and bred and maintained for 2 generations at the Veterans Affairs (VA) Boston Healthcare System animal resource facilities. Mice were genotyped using primers as indicated by Jackson Laboratories. Mice were housed individually and water and food were provided ad libitum throughout the experiments. Mice were maintained on a 12:12 h light/dark cycle [light onset Zeitgeber (ZT) 0] at 22 ± 3 °C. All experimental protocols were approved by the VA Boston Healthcare system Institutional Animal Care and Use Committee and were in compliance with the National Institutes of Health guidelines.

Nox2 knockout (-KO) mice (B6.129S-Cybb^tm1Din/J), Nox2^y/− (male) and Nox2^−/− (female), CD45.1 (B6.SJL-Ptprc^aPepc^b/BoyJ) and C57Bl/6J wild-type (WT) control mice were obtained from The Jackson Laboratory. In experiments using congenic system (CD45.1 vs. CD45.2), CD45.1 mice were used as Nox2^+/+ controls. Lyn^−/− mice (61 (link)) were backcrossed to a C57Bl/6 background as described (62 (link)). Akt1^−/− mice were generated in the laboratory of Nissim Hay as described (63 (link)). For NFκB reporter mice, we used a line of transgenic mice (HLL [HIV-LTR/Luciferase] mice) that carries the proximal 5’ human immunodeficiency virus (HIV-1) long terminal repeat (LTR), a well-characterrized NF-κB promoter, driving the expression of Photinus luciferase (64 (link)). HLL mice were crossbred with Nox2^−/− mice to generate Nox2^y/−-HLL mice. All the mice were bred and/or maintained in specific pathogen-free facilities at the University of Illinois at Chicago. For surgical procedures, anesthesia and perioperative analgesia were achieved with isoflurane and subcutaneous injection of buprenorphine SR LAB, respectively. Hindlimb ischemia surgery was performed as described previously (17 (link), 22 (link)). All procedures involving animals were approved by The Animal Care Committee at the University of Illinois at Chicago and conducted in accordance with institutional guidelines.