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6 µM rabbit GAPDH (Sigma-Aldrich) was incubated for 20 min in a total volume of 500 µl of 100 mM Tris/HCl p H 8.0, 1 mM EDTA, 1.4 mM H 2 O 2 and 3 mM GSH, which led to glutathionylation and inactivation of enzymatic activity

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Under oxidative stress, ROS act as major metabolic signals that enhance FoxO activation through multiple pathways: suppression of PI3K/Akt, activation of AMPK, JNK, and p38 MAPK, and deacetylation via SIRT1 and HDACs (Peris-Moreno et al., 2021)