In vivo treatment with the NFAT blocker A-285222 reduces plaque size in the brachiocephalic artery of old female IGF-II/LDLR–/–ApoB100/100 mice. (a) Histologically determined plaque size and (b) stenosis in the brachiocephalic artery of old mice treated with A-285222 (0.29 mg/kg) or vehicle (saline) for 4 weeks according to the same protocol detailed in Figure 1(a). N = 3 males, N = 4–7 females; *p < 0.05. (c) and (d) Plaque size determined non-invasively by ultrasound biomicroscopy before (baseline, week 4 of the experiment) and after treatment (week 8 of the experiment; N = 7–10 mice/group). (d) Change in plaque size from week 4 to week 8 for each group. (e) Young IGF-II/LDLR–/–ApoB100/100 mice had higher proportion of macrophage infiltration in subvalvular aortic plaques when compared to old mice (N = 4–6 young mice, N = 7–10 old mice). (f) Representative MOMA-2 stained sections of the subvalvular aorta from untreated young and old mice. Scale bar = 400 µm. (g) and (h) Gene expression of oxidative stress targets NOX4, NOX2, catalase and GLO1 (g) and inflammatory targets OPN, IL6, ICAM-1 and TF (h) in the aorta of old mice (N = 2–7 mice/group and sex). HPRT and β-actin were used as endogenous controls. *p < 0.05; **p < 0.01; and ***p < 0.001.