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For overexpression studies, pLX304-NINJ1 cDNA with a V5 tag was sourced from DNASU (HsCD00436123 and HsCD00438752), pWPI-SLC7A11 cDNA with an HA tag from Addgene (#201643), and pCDH-TFRC-mApple cDNA from Addgene (#179384)

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2 Methods 2.1 Animals and procedures The animal experimental procedures were approved by the Animal Care and Use Committee of Oujiang Laboratory (SYXK2023-0047, OJLAB22121609), and were carried out in strict accordance with the National Institutes of Health Guidelines for the Use of Laboratory Animals

Enzymatic deficiency of PAG leads to cerebral glutamine accumulation, which translates into dysfunctional excitatory signaling and associated encephalopathies (Rumping et al