Mitochondria were freshly isolated from brain tissues and washed two times with MAS. Mitochondrial pellets or neurosphere cells derived from cortices were lysed in buffer [50 mM tris-HCl (pH 7.4), 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA, 1 mM NaF, 1 mM Na3VO4, 1 mM phenylmethylsulfonyl fluoride, and aprotinin/leupeptin (10 μg/ml)]. Lysates were subjected to the PDH assay, pyruvate, acetyl-CoA, and α-ketoglutarate measurement using PDH activity colorimetric assay, pyruvate assay, acetyl-CoA assay, and α-ketoglutarate assay kits purchased from BioVision, following the manufacturers’ instructions.

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