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At 48 h post-transfection, cells were harvested in ice-cold PBS using Trypsin-EDTA (Lonza, Switzerland), centrifuged at 1000 g for 3 min, and washed with ice-cold PBS

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mTOR regulates differentiation and formation of OCs OCs, differentiated giant multinucleated cells derived from the monocyte/macrophage lineage, decompose the bone matrix by producing various enzymes and acids [107]