Nox2 Knockout Mice for Hindlimb Ischemia Study

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.