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Notably, acetylcarnitine can act as an acetyl donor [37], supporting the acetylation process of cells through the shuttle mechanism of acetylcarnitine in the absence of conventional acetyl-CoA synthetase enzymes such as ATP-citrate lyase (ACLY) and acyl-CoA synthetase short chain 2 (ACSS2) [38]

On the subsequent day, the membranes were washed three times with TBST buffer (CBW0089, COBIO) for 5 min each at room temperature, and afterwards, exposed to HRP-conjugated secondary antibodies (diluted 1:5000) for 30 min at room temperature

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Follow proper layering (thin to thick) rules and respect pH differences to avoid unnecessary over-concentration