Figure 4

Superoxide- and ROS-inhibitors reduce NETs in vivo. WT and TLR-4 KO mice underwent either sham laparotomy or 1 h of ischemia and 6 h of reperfusion, and received control, N-acetyl-cysteine (NAC; 150 mg/kg) or allopurinol (50 mg/kg) treatment. (A) ELISA assay of circulating in vivo NET MPO-DNA complexes present in the serum of WT mice, or (B) TLR-4 KO mice after I/R and treatment as above. Shown as mean fold over control. *p < .05 vs respective uninhibited treatment. (C) Western blot of in vivo citrullinated Histone H3 levels after sham or I/R and treatment as above from WT liver protein lysates, or (D) WT and TLR-4 KO after I/R and treatment as above. β-actin is shown as a loading control. (E) Assay of serum ALT levels in WT mice, or (F) TLR-4 KO mice after I/R and treatment as above. Data represent the mean ± SEM (n = 6 or more mice per group). *p < 0.05 vs sham. #p < .05 vs untreated WT I/R group. (G) Quantification of necrotic hepatocytes in H&amp;E stained liver sections from control-, NAC- or allopurinol-treated mice 6 h after reperfusion. *p < 0.05 vs WT control group after liver I/R; #p < .05 vs respective WT inhibitor-treated group after liver I/R.