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These combined mechanisms create a comprehensive metabolic effect that extends beyond glucose lowering

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By overexpressing tight junction proteins and maintaining the blood-brain barrier (BBB) integrity, short-chain fatty acids (SCFA) can penetrate the BBB via monocarboxylate transporters (MCTs) (67)

The cells were then washed three times with PBS containing 1% BSA (w/v) followed by 1 h incubation with 1:1,000 anti-mouse Alexa Fluor 488 conjugated secondary antibody (diluted with PBS containing 5% BSA, Invitrogen) at RT in the dark