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To test the hypothesis that relatively few amino acid residues determine substrate selectivity, the Y108V mutant of GSTP was studied (Nuccetelli et al

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PaSTRy Solutions of EV-F3 and additive at either 100 or 1000 molar excesses (corresponding to 0.1 mM or 1 mM final concentration) were aliquoted into a 96-well PCR plate containing SYTO9 dye (Thermo Fisher Scientific) in triplicate with 50 mM HEPES, pH 8.0, 200 mM NaCl buffer, before centrifugation for 3 min at 4 C

For each assay, cells were plated in duplicate, with one plate containing CellTox Green Dye added according to the Express, No-Step Addition at Seeding protocol described in the CellTox Green Cytotoxicity Assay Technical Manual #TM375 (10l of CellTox Green Dye per 5ml of cells)