Neutrophil depletion experiment in ischemia reperfusion mouse model

To elucidate the role of neutrophils in the molecular alterations observed after I/R, we performed an experiment of I/R in neutrophil-depleted mice. Neutrophil depletion was performed by anti-Ly6G injection and IgG was used as vehicle. We have performed an unbiased, open-search analysis of the oxidative PTMs and quantitative multiplexed proteomics that take place in the mouse heart tissue at 24h after I/R. Neutrophil depletion was performed in 3 mice by Ly6G injection 24h and 48 h prior to I/R and sacrificed 24 hours after reperfusion. A second group consisting on 3 mice that were injected with IgG antibody as vehicle, and subjected to I/R and sacrificed 24h after reperfusion. A third group of 3 mice with no injection were also sacrificed 24h after reperfusion. 12 additional mice from the three previous groups were sacrificed without I/R intervention: control without injection (n=6), IgG injection (n=3) and Ly6G injection (n=3). Whole hearts tissue was collected for proteomics evaluation.