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The qPCR reaction was 95°C for 10 minutes, 40 cycles of denaturation at 95°C for 15 seconds, and a combined annealing/extension at 60°C for one minute

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Senescence-associated β-galactosidase (SA-β-gal) activity was measured in both cells and frozen tissues using a commercial kit, providing a visual and quantitative indicator of senescent cells

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