Western blot protocols
Protein extracts from skeletal muscle (4 μg for PRX2/3) and erythrocytes (6 μg for PRX2) were separated by SDS-PAGE under non-reducing conditions. Extracts from skeletal muscle (30 μg for NRF2, GPx1, SOD1 or 40 μg for Akt, AS160, and PAK1/2) were separated by SDS-PAGE under reducing conditions. Separated proteins were transferred to PVDF membranes, stained and imaged for total protein using Ponceau S (Sigma-Aldrich, Cat. P3504) prior to blocking for 1h in Tris-Buffered Saline/0.1%Tween-20 (TBS-T) containing 3% BSA. Blocked membranes were then incubated overnight at 4 °C with the following antibodies diluted in TBS-T/3%BSA; rabbit polyclonal anti-human PRX2 at 1:10,000 (R8656, Sigma-Aldrich, RRID: 