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For the extraction, methanol and internal standards were added, followed by incubation for 15 min at 30C on a thermal shaker (650 rpm, TS1 ThermoShaker, Biometra, Gttingen, Germany), centrifugation (21,000 x g, 10 min, 4C, Heraeus Fresco 21 Centrifuge, Thermo Scientific, Schwerte, Germany), evaporation, and centrifugation with methanol again

DOI: 10.1186/1743-7075-7-30 Inazu, M., & Matsumiya, T

In addition to ROS overproduction, the activation of MAPK and inactivation of phosphatidylinositol 3-kinase/Akt (a serine/threonine kinase that is also known as protein kinase B)/mammalian target of rapamycin (mTOR) are associated with both apoptosis and autophagy 6