Preparation of skeletal muscle tissue samples for biochemical and Western blot analyses
COLD
For GSH/GSSG and oxiPRX2/3, muscle tissue proteins were extracted as adapted from Kumar et al. [For Western blot analyses of NRF2, GPx1, SOD1, Akt, AS160, and PAK1/2, 30 mg of snap-frozen muscle tissue was homogenized in ice cold RIPA buffer (15 mM Tris, 167 mM NaCl, 0.5% sodium, deoxycholate, 0.1% SDS, 1% Triton X-100), containing complete protease and phosphatase inhibitor cocktail (at 25 μl per mg tissue, using an automatic homogenization blender (IKA) for 1 min. Sample lysates were placed in an orbital shaker for 1 h at 4 °C before centrifuging at 600×