Fig. 3

Pulmonary NO production and eNOS phosphorylation in orthotopic 4T1 breast cancer model in mice. a NO production in lungs, presented as mean ± SD. Data were Box–Cox transformed and analyzed with one-way ANOVA followed by Tukey’s multiple comparison test; n = 10 for control and 1st–4th week of disease; n = 19 for 5th week of disease. Black circle indicates outlier. Perfused lungs were excised from euthanized animals and NO production measured (see Methods). Statistical significance vs healthy control mice at **P < 0.01 and ***P < 0.001. b Densitometric data presenting eNOS and p(S1177)eNOS levels during progression of breast cancer used to calculate relative eNOS phosphorylation at serine 1177 (black circle) expressed as fraction of total eNOS level in lungs of control healthy mice for which an arbitrary value of 1 was ascribed. Western blot image presents fold change vs control sample corresponding to healthy mice. Total protein after transfer was used as loading control. Western blot image shows p(S1177)eNOS and eNOS levels in pooled samples in all experimental groups obtained by pooling lung homogenates from six mice in each experimental group (see Methods). AU arbitrary units, eNOS endothelial nitric oxide synthase, NO nitric oxide