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Briefly, buffers provided with the kit were supplemented with Protease Inhibitor Cocktail (P8340, Sigma) and used to lyse the cells with a Dounce homogenizer, followed by separation by variable speed centrifugation

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However, as a NAD-dependent deacetylase that modulates apoptosis of hepatocytes in response to oxidative and genotoxic stress, SIRT1 inactivation could provoke insulin resistance and pro-inflammatory cytokine (TNF-, IL-1, and IL-6) expression and pro-inflammatory signaling pathway (STAT3 and JNK) activation 135 , all of which are important factors governing hepatoma cells transformation

p53 also promotes autophagy by upregulating negative regulators of the AKT/mTOR pathway, such as PTEN and sestrins [81]

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Similarly, upon DNA single-strand breaks (SSBs), p53 is activated by the ataxia telangiectasia mutated- and Rad3-related kinase (ATR)/p53 pathway