Anti sod2

The following antibodies were used in the present study: anti-SOD2 from Enzo Life Sciences (Farmingdale, NY, USA); anti-IL-6 and anti-β-catenin from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-Bcl-X_L, anti-STAT3, anti-Src, and anti-phospho-Src from Cell Signaling Technology (Danvers, MA, USA); and anti-β-actin from Sigma-Aldrich (St. Louis, MO, USA). Recombinant human IL-6 was purchased from Millipore (Darmstadt, Germany).

PKC epsilon, SOD2, Nrf-2, and Myogenin protein content was tested with a Western blot. In brief, C2C12 was lysed with a RIPA buffer and total protein was quantified with a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA), according to the instruction protocol. Forty micrograms of proteins were separated on 10% SDS-polyacrylamide gel electrophoresis, transferred to a nitrocellulose membrane, and incubated with a specific primary antibody diluted in the blotting solution as recommended by the manufacturer protocol. We used anti-PKCe (Merck Millipore, Darmstadt, Germany, Cat. No.: 06-991), anti-SOD2 (Enzo Life Sciences Inc., Farmingdale, NY, USA, Cat. No.: ADI-SOD-111F), anti-Myogenin (Santa Cruz, Dallas, TE, USA, Cat. No.: sc-12732), anti-Nrf2 (clone: D179C, Cell Signaling, Danvers, MA, USA, Cat. No.: 12721), and anti-HSP70 (Sigma-Aldrich, St. Louis, MO, USA, Cat. No.: H5147) as a loading control. The nitrocellulose membranes were then washed and incubated with 1:5000 peroxidase-conjugated anti-rabbit (Thermo Fisher Scientific, Waltham, MA, USA) or 1:2000 peroxidase-conjugated anti-mouse IgG (Sigma-Aldrich, St. Louis, MO, USA). Proteins were resolved with an ECL SuperSignal West Pico Chemiluminescent Substrate Detection System (Thermo Fisher Scientific, Waltham, MA, USA) and densitometric analyses were performed using ImageJ software (by NIH).

The detailed procedure has been described^{24 (link)}. The following primary antibodies were used: anti-Fxn (GeneTex, Cat.#: GTX54036), anti-Cox4 (Cell Signaling Technology, Cat.#: 48445), total OXPHOS Rodent WB Antibody Cocktail (MitoSciences, Cat.#: MS604), anti-Sod1 (Abcam, Cat.#: ab16831), anti-Sod2 (Enzo Life Sciences, Cat.#: ADI-SOD-111-D), anti-Sod3 (R&D Systems, Cat.#: AF4817), anti-Cat (Abcam, Cat.#: ab15834), anti-β-Tublin (Cell Signaling Technology, Cat.#: 2146 S), anti-Mmp9 (Proteintech, Cat.#: 10375-2-AP), anti-Col1a2 (Proteintech, Cat.#: 14695-1-AP), anti-Irp1 (Santa Cruz Biotechnology, Cat.#: sc-166022), anti-Tfrc (Abcam, Cat.#: ab84036), anti-Ft (Abcam, Cat.#: ab75973), anti-Opa1 (Novus Biological, Cat.#: NBP2-34206), anti-4-HNE (Abcam, Cat.#: ab48506) and anti-Gapdh (Cell Signaling Technology, Cat.#: 14C10). The proteins were detected and quantified using an Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE).