Fig. 6.

Changes in markers of oxidative stress and fibrosis after chronic activation of DAAO. a Relative changes in expression of the Nrf2 targets Hmox1, Nqo1, Sxn1, and Txnrd1 and the NF-κB targets Il1b, Tnfa, Icam1, and Nos2 in hearts from animals infected with control AAV9 (Ctrl) and DAAO virus measured by qPCR. Distributions for animals infected with control virus can be found on Supplementary Figure 4C. **p < 0.01 and ***p < 0.001 by ANOVA. b Relative changes in expression of the reductive enzymes Prdx1, Prdx2, Prdx3, Gpx1, Gpx3, Txn1, and Txn2 in Ctrl and DAAO hearts measured by qPCR. Distributions for animals infected with control virus can be found on Supplementary Figure 4D. c Reduced (GSH) measured in hearts from animals expressing DAAO vs. Ctrl. **p < 0.01 by t-test. d Total glutathione measured in hearts from DAAO-expressing vs. control animals treated with D-alanine for 4 weeks. *p < 0.05 by t-test. e Immunoblot and densitometry for the fibrotic marker galectin-3 in cardiac lysates from Ctrl and DAAO animals. f Relative changes in expression of the fibrosis-associated transcripts Col1a1, Col3a1, Tgfb1, and Mmp2 in hearts from Ctrl (blue circles) and DAAO (red squares) animals. No significant differences were observed by ANOVA. g Representative histology of hearts from Ctrl and DAAO animals stained for collagen with Masson’s trichrome stain. h Masson trichrome-stained cardiac tissue sections quantitatively analyzed for fractional area of fibrosis. The images shown are representative of n = 4 animals from each group, which were pooled for statistical analysis. Data are represented as mean ± standard error