Figure 11.

Association of Nrf2 protein with mitochondria. Neonatal rat cardiomyocytes with or without Nrf2 adenoviral transfection were treated with 300 μM H₂O₂ for 2 h. Cells were immediately harvested and fractionated into cytosolic, mitochondrial, and nuclear fractions for Western blot analysis of Nrf2 and Keap-1, along with protein markers of each fraction: GAPDH (cytosol), ND2 (mitochondria), and TBP (nucleus) (A). An equal amount of proteins from cytosolic, mitochondrial, or nuclear fractions isolated from rat heart tissues were used for Western blot to detect for Nrf2 and Keap-1 with protein markers (B). Mitochondria isolated from rat liver were subfractionated into whole mitochondria, mitoplast, and outer mitochondria membrane as described in Materials and Methods for Western blot analysis of Nrf2 and protein markers of each fraction: GAPDH (cytosol), OPA1 (mitoplasts), and Bcl-xL (OMM) (C).