At the designated time points, 3T3-L1 cells were washed with PBS and homogenized in a modified RIPA buffer (Sigma, St. Louis, MO, USA) containing a proteinase inhibitor cocktail. Later, lysates were collected by centrifugation at 12,074× g for 20 min at 4 °C. Protein concentration was evaluated by bicinchoninic acid (BCA) protein assay kit (Thermo Scientific, Rockford, IL, USA) [49 (link)].
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