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. Author manuscript; available in PMC: 2019 Jan 5.
Published in final edited form as: Circ Res. 2017 Nov 1;122(1):58–73. doi: 10.1161/CIRCRESAHA.117.311307

Figure 2. Mitochondrial respiratory function and ROS production in isolated mitochondria from ACStg hearts.

Figure 2

(A,B) Maximal ADP-stimulated mitochondrial oxygen consumption (A) and ATP synthesis rates (B) in mitochondria isolated from 12- and 24-week-old WT and ACStg hearts using palmitoyl-carnitine as a substrate; n =4-5. * P<0.05 vs. WT. (C,D) Maximal ADP-stimulated mitochondrial oxygen consumption in saponin-permeabilized cardiac fibers of 12- and 24-week-old WT and ACStg hearts using pyruvate (C) or glutamate (D) as a substrate; n =6. (E-G) Electrophoretic separation of OXPHOS complexes by blue-native PAGE (E), representative images of in-gel activities of complexes I, IV, and V (F), and quantification of OXPHOS complex activities (G), measured in 12-week-old WT and ACStg hearts; n =4. *P<0.05 vs. WT. (H) H2O2 production with succinate as a substrate in mitochondria isolated from 12- or 24- week-old WT and ACStg hearts, or with palmitoyl-carnitine as a substrate in mitochondria from 12-week-old WT and ACStg hearts, in the absence or presence of rotenone (Rot); n =3 at 12wks, n =6 at 24wks. *P<0.05 vs. WT. (I) Oxidation of DCFDA in whole tissue extracts of 24-week-old WT and ACStg hearts; n =5-6. *P<0.05 vs. WT. (J) Western blot for 4HNE protein adducts in mitochondrial protein isolated from WT and ACStg hearts. Numbers beneath each lane represent densitometry of 4HNE immunoreactivity of all bands in that lane. (K) Aconitase activity measured in mitochondria isolated from 12- or 24-week-old WT and ACStg hearts; n =3 at 12weeks, n =6 at 24weeks. All data are mean ± sem.