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Brain tissue (100 mg) was taken in 900 microliters of PBS (1:9) ratio in a glass homogenizer on ice and centrifuged at 2,000 rpm for 5 minutes

The BBB employs various transport systems, such as carrier-mediated transport (CMT), receptor-mediated transport (RMT), active efflux transport, vesicular trafficking by transcytosis, and ion transport, to facilitate the entry of essential substances into the brain parenchyma and the efflux of metabolic waste and endogenous neurotoxins [19, 23]

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Harvested supernatants were used for gene knockout