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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: Arthritis Rheumatol. 2014 Sep;66(9):2532–2544. doi: 10.1002/art.38703

Figure 4. NET-derived NOS and NOX induce HDL oxidation in mice, while NET inhibition decreases HDL oxidation in vivo.

Figure 4

Immunofluorescent staining and immunoblotting was used to identify possible enzymatic sources of HDL nitration. (A) NOX subunits, p22 and p47, and two NOS isoforms, eNOS and iNOS, were detected in murine NETs. Control Balb/c and lupus NZM NETs were added to control, unoxidized murine HDL. (B) Resulting levels of MPO-specific Cl-Tyr oxidation in the absence or presence of isolated NETs, as quantified by mass spectrometry (N=4/group). (C) Resulting levels of NOS- and NOX-mediated N-Tyr oxidation as quantified by mass spectrometry (N=8/group). 3-AT, NMMA and DPI were added to block MPO, NOS and NOX activity, respectively. Absence of PMA stimulation was used as a negative control for NET formation. (D) Levels of plasma and HDL N-Tyr content in 26-week old NZM mice that received daily injections of PBS (N= 10) or Cl-Am (N=10) for 14 weeks. (For C, comparisons were made within the same mouse strain; ** p<0.01, **** p<0.0001. Data are displayed as mean ± SEM).