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For NanoLuc assay measurements, the homogenate (10 L) was diluted in reporter lysis buffer (40 L) and sample (50 L) was further diluted in NanoLuc substrate (50 L) [Nano-Glo luciferase assay, 5000x dilution from stock, Promega Cat#N1110] and the luminescence were measured through an open filter

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In such severe cases, cerebral oedema becomes clinically apparent 12 hours to 4 days after the overdose [18, 20, 50, 87], although CNS depression may be delayed if a slow-release preparation has been ingested [12, 50]