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Online Resource Role of endothelial Nox2 NADPH oxidase in angiotensin II-induced hypertension and vasomotor dysfunction. Colin E Murdoch, Sara P Alom-Ruiz, Minshu Wang, Min Zhang, Simon Walker, Bin Yu, Alison Brewer, Ajay M Shah Basic Research in Cardiology

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Online Resource

Role of endothelial Nox2 NADPH oxidase in angiotensin II-induced hypertension and vasomotor dysfunction.

Colin E Murdoch, Sara P Alom-Ruiz, Minshu Wang, Min Zhang, Simon Walker, Bin Yu, Alison Brewer, Ajay M Shah

Basic Research in Cardiology

King’s College London British Heart Foundation Centre, Cardiovascular Division, London

SE5 9PJ, UK.

Corresponding author ajay.shah@kcl.ac.uk

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Online Resource 1:

Comparison of body and organ weight from Nox2Tg and WT littermate controls.

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A

C

B

Tg

WT

Nox2

eNOS

Actin

actin

*

Online Resource 2: (A)Nox2 protein levels in aorta from line 2 Nox2 Tg mice (*= P<0.05; t-test; n=4). (B) NADPH-dependent superoxide production assessed by lucigenin-enhanced chemiluminescence in aortic homogenate. Representative bar graph showing reduction in chemiluinescence signal after pre-treatment (15min) with tiron, DPI or apocynin (Apo), whereas pre-treatment with L-NAME, rotenone or, allopurinol (Allo) had no effect on the superoxide production (* p<0.05, **p<0.01; n=6; One-way ANOVA, Bonferroni post-hoc test). (C) Representative blots showing protein expression of eNOS and actin in Wt and Nox2 Tg aorta with the corresponding mean data ( P=NS; n>3).

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Online Resource 3: Effect of in vivo angiotensin II treatment on aortic antioxidant mRNA levels. WT and Tg groups were treated with either saline or AngII (1.1mg/kg/day). There were no differences between Wt and Tg in the response to AngII by 2-way ANOVA. Asterisks indicate significant effect (p<0.05) of AngII treatment within Wt or Tg groups. N>3 per group.