Manganese superoxide dismutase mnsod

Total proteins were extracted from kidneys using a lysis buffer. Extracted proteins were separated on gradient sodium dodecyl sulfate polyacrylamide gels and transferred to nitrocellulose membranes. The membranes were probed with primary antibodies against NGAL (Santa Cruz Biotechnology, Santa Cruz, CA, USA), kidney injury molecule-1 (KIM-1; Abcam, Cambridge, MA, USA), c-Jun N-terminal kinase (JNK; Cell Signaling Technology, Danvers, MA, USA), p-JNK (Cell Signaling Technology, Danvers, MA, USA), extracellular signal-regulated kinase (ERK; Cell Signaling Technology, Danvers, MA, USA), p-ERK (Cell Signaling Technology, Danvers, MA, USA), p38 (Cell Signaling Technology, Danvers, MA, USA), p-p38 (Cell Signaling Technology, Danvers, MA, USA), NF-κB p65 (Cell Signaling Technology, Danvers, MA, USA), p-NF-κB p65 (Cell Signaling Technology, Danvers, MA, USA), NADPH oxidase 4 (NOX4; Novus Biologicals, Littleton, CO, USA), manganese superoxide dismutase (MnSOD; Abcam, Cambridge, MA, USA), p53 (Cell Signaling Technology, Danvers, MA, USA), Bax (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Cell Signaling Technology, Danvers, MA, USA). The signals were detected using chemiluminescence detection reagents (Thermo Fisher Scientific, Waltham, MA, USA).

Tissues were lysed on ice in a RIPA lysis buffer. The extracted proteins were loaded onto precast gradient polyacrylamide gels and then transferred to nitrocellulose membranes. The membranes were reacted with antibodies against α-SMA (Sigma-Aldrich, St. Louis, MO, USA), NADPH oxidase 4 (NOX4; Novus Biologicals, Littleton, CO, USA), catalase (Abcam, Cambridge, MA, USA), manganese superoxide dismutase (MnSOD; Abcam, Cambridge, MA, USA), cleaved caspase-3 (Cell Signaling Technology, Danvers, MA, USA), cleaved poly(ADP-ribose) polymerase-1 (cleaved PARP-1; Cell Signaling Technology, Danvers, MA, USA), Bax (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-IκB-α (Cell Signaling Technology, Danvers, MA, USA), IκB-α (Cell Signaling Technology, Danvers, MA, USA), p-NF-κB p65 (Cell Signaling Technology, Danvers, MA, USA), NF-κB p65 (Cell Signaling Technology, Danvers, MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Cell Signaling Technology, Danvers, MA, USA). Then, the membranes were probed with secondary antibodies. The bands were visualized on the iBright CL1500 Imaging System (Thermo Fisher Scientific, Waltham, MA, USA) using enhanced chemiluminescence reagents. GAPDH was used as a loading control.