Immunohistochemical Analysis of Cellular Markers

Immunohistochemistry was performed as described previously [27 (link)]. The following primary antibodies were used: monoclonal anti-CAV-1 (1:250; Epitomics, Burlingame, CA, USA), monoclonal CyP A (1:150; GeneTex, Irvine, CA, USA), monoclonal anti-peroxisome proliferator-activated receptor-γcoactive 1α (PGC-1α; 1:250; Abcam), polyclonal anti-nuclear respiratory factor-1 (NRF-1; 3 μg/mL; GeneTex), polyclonal anti-superoxide dismutase 2 (SOD2; 1:1000, Novus, Littleton, CO, USA), and polyclonal anti-catalase (1:1000; Abcam). monoclonal anti-nuclear factor E2-related factor 2 (Nrf2; 1:250; GeneTex), polyclonal anti-Kelch-like ECH- associated protein 1 (Keap1; 1:150; Bioss, Woburn, MA, USA), and polyclonal anti- glutamate-cysteine ligase catalytic subunit (GCLC; 1:100; GeneTex). Nuclei were counterstained with hematoxylin and photographed using an Olympus BX61 microscope (Tokyo, Japan). For animal kidney tissue, positive cells were quantified in five fields per animal at 400×, with six rabbits for each group per time point (Image-Pro Plus 4.5).