Figure 3.

Trx1 is required for redox regulation in the heart. (A) Expression levels of reducing enzymes in Trx1cKO mice. Expression levels of indicated reducing enzymes were examined with RNA-seq. The mean value from wild-type mice was expressed as 1 (N = 3). (B) Increased monomer form of Prdx1 in response to H₂O₂ in primary cultured cardiomyocytes. Cardiomyocytes were incubated with 100 μM H₂O₂ for 30 min. Cell lysates were resolved by SDS-PAGE under non-reducing conditions and subjected to western blot analysis. (C) Increased monomer/dimer ratio of Prdx1 in Trx1cKO mice. Heart lysates from Trx1cKO mice were subjected to SDS-PAGE under non-reducing conditions and subsequent western blot analysis. Monomer and dimer signal intensities were quantified (Right). (D) Increased sulphonic acid formation in Prdx1 in Trx1cKO mice. (E) Decrease in reduced protein thiols in Trx1cKO mice. Reduced protein thiols were labelled with biotin-IAM (iodoacetamide). The labelled proteins were visualized with HRP-streptavidin. (F–G) Increased oxidative protein adducts such as 4HNE (4-hydroxynonenal) (F) and DiTyrosine (G). N5–6 (C and D) and 6 (E–G). Statistical significance was determined with the Student’s t test (A–G). *P<0.05, **P<0.01, and ***P<0.001.