MnSOD and bcl-xL mRNA expression are not changed following treatment with ADR.

<p>Real time PCR was used to evaluate (a) MnSOD and (b) Bcl-xL mRNA levels (6 h) following ADR treatment in WT, iNOS (−/−), TgM (++), and iNOS (−/−)-TgM (++) mice.</p

NFκB DNA binding activity is increased in WT mice.

<p>Nuclear extract (12 µg) from heart tissue of WT, iNOS (−/−), TgM (++), and iNOS (−/−)-TgM (++) mice was isolated and assayed for NFκB DNA binding activity. Densitometric quantitation as fold increase from respective saline control is shown (with gel shift inset). *p<0.001 versus saline for WT.</p

Serum nitrate is significantly increased in WT but not iNOS (−/−), TgM (+/+), or iNOS (−/−)-TgM (+/+) mice following treatment with ADR.

<p>Serum nitrate concentration (µM) following time course treatment with ADR (n = 6–8 mice for each time point) was determined. Values are expressed as mean ± SEM. *p<0.0001 versus saline for WT mice.</p

p53 DNA binding activity is significantly increased in WT and mice lacking iNOS.

<p>Nuclear extract (12 µg) from heart tissue was isolated, pooled (N = 4–6 animals per lane) from (a) WT and iNOS (−/−), (b) TgM (++), and (c) iNOS (−/−)-TgM (++) mice, and assayed for p53 DNA binding activity. Specificity of DNA binding activity was determined using an anti-p53 antibody, as well as competition with unlabeled p53 oligonucleotide. Densitometric quantitation as fold increase from respective saline control using both individual animals and pooled samples is shown in (d). *p<0.025 versus saline for WT and iNOS (−/−) mice.</p

Induction and interaction of p65 with p53 in nuclear extract following treatment with ADR.

<p>Expression of p65 increases following treatment with ADR (A). Immunoprecipitation with p65 demonstrating an interaction with p53 in the nucleus after treatment with ADR. Anti-p65 was used to reprobe membrane to verify presence of p65 (B).</p

Transgenic mice overexpressing MnSOD have increased enzyme activity and protein expression.

<p>(a) Superoxide dismutase activity of heart tissue homogenate in four genotypes: WT, iNOS (−/−), TgM (++), and iNOS (−/−)-TgM (++). Data are expressed as mean ± SEM. *p<0.05 for MnSOD compared to WT and iNOS (−/−). (b) Protein extracts were analyzed by Western blot using an antibody detecting MnSOD. Protein extracts were co-hybridized with an antibody detecting CuZnSOD to assess equal loading of the samples.</p

Bax mRNA expression is significantly increased in iNOS (−/−) mice following treatment with ADR.

<p>Real time PCR was used to evaluate Bax mRNA expression (6 h) following ADR treatment in WT, iNOS (−/−), TgM (++), and iNOS (−/−)-TgM (++) mice. Data are expressed as fold increase from respective saline control. <b>*</b>p<0.025 versus saline for iNOS (−/−) mice.</p