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The tissue samples were homogenised in phosphate-buffered saline (PBS, pH 7.4) containing 0.1 mM butylated hydroxytoluene (BHT) (TissueRuptor, Qiagen, Valencia, CA, USA) and sonicated for 30 s in an ultrasonic ice bath (Branson, USA)
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The results showed that after FOXO4-DRI treatment, the p-P53/P53 ratio significantly increased in the nucleus, cytoplasm, and total protein samples (Figure 5D)