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Considering the abundance of Hb in RBCs (corresponding to 10 m M heme), the reaction of autoxidation of Hb is therefore the most abundant source of ROS in RBCs

Agents such as metformin, rapamycin, or trehalose can rejuvenate autophagy and mitophagy, thereby clearing damaged mitochondria and lipid droplets to reduce mitochondrial ROS and inflammatory priming

Medesan C, Cianga P, Mummert M, Stanescu D, Ghetie V, Ward ES

Briefly, protein extracts were prepared by homogenizing with a Polytron homogenizer (KINEMATICA AG, Switzerland) in lysis buffer containing: 20 mM HEPES (pH 7.9), 25% v/v glycerol, 0.42 M NaCl, 0.2 mM EDTA, 1.5 mM MgCl2, 0.5% v/v Nonidet P-40, 1 mM DTT, 1 mM Naf, 1 mM Na 3 VO 4 , and 1 complete protease inhibitor cocktail (Roche Diagnostics Ltd, Mannheim, Germany)