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Cellular viability Optimization of Sp concentration and its cytotoxicity parameters were established by treating of 1.5 10 5 HeLa or 1.2 10 5 SHSY-5Y cells with serial concentrations of the Sp (100 M1 mM) for 15 min or conducting a time-course (5 min48 h) using 250 M Sp, followed by MTT assay as per standard protocol (Thermo Fisher Scientific)

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R162A and S353Y double mutant) were used to replace the EGFP gene, generating the AAV-hHPN WT and AAV-hHPN RS vectors