Extended Data Figure 8. iNOS overexpression in cardiomyocytes reduces IRE1α activation and Xbp1s levels without affecting cardiomyocyte viability.

a, Immunoblot images of iNOS and GAPDH proteins of neonatal rat ventricular myocytes infected with increasing multiplicity of infection (MOI) of α-galactosidase adenovirus (AdLacZ) or iNOS adenovirus (AdiNOS) for 24 hours. Images are representative of three independently performed experiments with similar results. b, iNOS mRNA levels in neonatal rat ventricular myocytes transduced with increasing MOI of AdlacZ or AdiNOS for 24 hours (n=4 biologically independent experiments). c, Medium nitrite/nitrate concentration of neonatal rat ventricular myocytes transduced with increasing MOI of AdlacZ or AdiNOS for 24 hours (n=4 biologically independent experiments). d, Lactate dehydrogenase (LDH) release in neonatal rat ventricular myocytes transduced with increasing MOI of AdlacZ or AdiNOS for 24 hours (n=3 biologically independent experiments). e, Immunoblot images of Cys-SNO, iNOS and GAPDH proteins in NRVMs transduced with AdlacZ or AdiNOS for 24 hours (100 MOI). Images are representative of three independently performed experiments with similar results. f, Immunoblot images of pIRE1α, IRE1α, iNOS and GAPDH proteins in neonatal rat ventricular myocytes transduced with increasing MOI of AdlacZ or AdiNOS in the presence or absence of tunicamycin (TUN) for 24 hours. Images are representative of three independently performed experiments with similar results. g, Xbp1s mRNA level of neonatal rat ventricular myocytes transduced with 100 MOI of AdlacZ or AdiNOS in the presence or absence of TUN for 24 hours (n=3 biologically independent experiments). Results are presented as mean±S.E.M. b-d, g One-way ANOVA followed by Sidak’s multiple comparisons test. Numbers above square brackets show significant P values. For gel source data, see Supplementary Fig. 1.