Supplementary Table 1. Table showing different gene specific primers used in real-time PCR.

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Supplementary Table 1. Table showing different gene specific primers used in real-time PCR. gene Forward (5 3 ) Reverse(5 3 ) act CGTGAAAAGATGACCCAGATCA TGGTACGACCAGAGGCATACAG Nox1 TCGACACACAGGAATCAGGA TTACACGAGAGAAATTCTTGGG Nox2 GGTTCCAGTGCGTGTTGCT GCGGTGTGCAGTGCTATCAT Nox2 (KO) ACTCCTTGGGTCAGCACTGG GTTCCTGTCCAGTTGTCTTCG Nox3 CCGGCAGATCCAATAGAAGT GTCACTCCCTTCGCCTCTCT Nox4 GGAGACTGGACAGAACGATTCC TGTATAACTTAGGGTAATTTCTAGAGTGAATGA Duox1 GGCCTGGATTCTCCTGGTTGGGA AACAAGGCGCTGCAGCCGAG Duox2 CAGCGCTACGACGGCTGGTT GCTTAGCCGGCGAGGGTTGG P47 AGAGTCGCCAGGGCACTCT TCTTCGCCTGGCTGTCAGT P67 AAGCTGTTTGCCTGTGAGGT CTTCATGTTGGTTGCCAATG P22 GGAGCGATGTGGACAGAAGTA GCACCGACAACAGGAAGTG SOD1 GGACCTCATTTTAATCCTCACTCTAAG GGTCTCCAACATGCCTCTCTTC SOD2 CACACATTAACGCGCAGATCA GGTGGCGTTGAGATTGTTCA SOD3 GGGATGGATCTAGAGCATTAAGGA ACACCTTAGTTAACCCAGAAATCTTTTC enos CTGGAGCACCCCACGCT AGCGGTGAGGGTCACACAG catalase GCTGAGAAGCCTAAGAACGCAAT CCCTTCGCAGCCATGTG vecad TCAACGCATCTGTGCCAGAGAT CACGATTTGGTACAAGACAGTG

Supplementary Figure 1. (A) Contraction curves of aortic rings from ESMIRO mice with indicated acute treatments in response to phenylephrine (PE). (B) Contraction curves of gp91ds-tat treated aortic rings from ESMIRO mice before and after insulin (100mU/mL for 2 hours) treatment in response to PE. (C) Normalised oxidised DHE level in endothelium denuded aorta of wild type (WT) and ESMIRO mice measured by HPLC based DHE analysis (n=4 mice in each group).

Supplementary Figure 2. Increase in PE (10M) induced tension by L-NMMA (100M for 0.5 hours) in aortic rings from IR +/- mice with indicated acute treatment (n= at least 4 mice in each group * denotes P<0.05).

Supplementary Figure 3. (A) Relative Nox2 mrna expression in aorta, lungs and spleen of wild type (WT) and ESMIRO mice. (B) Relative Nox2 mrna expression in endothelium denuded aorta of WT and ESMIRO. (C) Relative vecad mrna expression in whole aorta and endothelium denuded aorta of WT and ESMIRO mice (n= at least 4 mice in each group; * denotes P<0.05).

Supplementary Figure 4. Normalised Nox2 protein expression in PEC of ESMIRO mice after Nox2 or scrambled sirna treatment (n=3; insets show representative blots of Nox2 and -actin * denotes P<0.05).

Supplementary Figure 5. Effect of chronic infusion of gp91ds-tat or scrambled peptide on weight and fasting insulin in ESMIRO or IR +/- mice. (A) Normalised weight of individual organs of ESMIRO. (B) Mean plasma insulin levels of ESMIRO. (C) Normalised weights of individual organs of IR +/-. (D) Mean plasma insulin levels in IR +/- mice (n=at least 4 mice in each group).

Supplementary Figure 6. Relative mrna expression of different ROS related genes (SOD=superoxide dismutase) in PEC of ESMIRO mice infused with gp91ds-tat or scrambled peptide through osmotic pump (n=3 mice in each group; * denotes P<0.05).

Supplementary Figure 7. (A) Relative expression of Nox2 mrna in lung and spleen of ESMIRO and ESMIRO/Nox2 y/- mice. (B) Normalised organ weight (C) blood pressure (D) plasma insulin concentration of ESMIRO and ESMIRO/Nox2 y/- mice (n= at least 4 mice in each group).

Supplementary Figure 8. Relative mrna expression of different ROS related genes (cat denotes catalase, SOD=superoxide dismutase) in PEC of ESMIRO and ESMIRO/Nox2 y/- mice (n=3 mice in each group; * denotes P<0.05).

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