T Cell-Derived IL-17A Induces Vascular Dysfunction via Perivascular Fibrosis Formation and Dysregulation of ^⋅NO/cGMP Signaling

Figure 3

No eNOS uncoupling in CD4-IL-17A^ind/+ mice. (a) NO production in aortic tissue was measured via electron paramagnetic resonance- (EPR-) based spin trapping. a2 shows the quantification of the delta intensity normalized to dry weight of samples; ; Student’s unpaired -test. (b) Oxidative fluorescence microtopography of aortic sections of CD4-IL-17A^ind/+ and IL-17A^ind/+ mice. b1: representative picture of isolated aortic sections stained with dihydroethidium (DHE, 1 μM). Autofluorescence of the laminae is visible in green, superoxide formation in red fluorescence; . b2: quantification of superoxide formation in the aortas is shown as a percentage of control; ; unpaired Student’s -test. (c) ROS formation in aortic tissue (endothelial scan) of CD4-IL-17A^ind/+ and IL-17A^ind/+ mice, incubated with L-NAME (eNOS inhibitor) or buffer. c1: representative DHE photomicrotopographs of aortic cryosections with focus on the endothelial layer; ROS formation appears in red. c2: quantification; ; comparison of IL-17A^ind/+ control mice with CD4-IL-17A^ind/+ mice or comparison of IL-17A^ind/++L-NAME mice with CD4-IL-17A^ind/++L-NAME mice; Mann–Whitney -test. (d) S-Glutathionylation (GSH) of eNOS was determined by eNOS immunoprecipitation from aortic protein samples of CD4-IL-17A^ind/+ and IL-17A^ind/+ mice, followed by antiglutathione staining and normalization to eNOS. Disappearance of the antiglutathione staining in the presence of 2-mercaptoethanol (2-Me) served as a control. d1: representative original blot. d2: densitometric analysis; (per , 3-4 aortas were pooled); Mann–Whitney -test. Data are shown as .

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