β-Galactosidase–expressing viruses HSV-1 gL86 and HSV-2 (333)gJ at an MOI of 10 were used in this study. Cells were plated at a density of 1 × 104 in 96-well plates overnight before use. Neutralization or prophylactic treatment using HPAC at concentrations of 1, 0.5, and 0.1 mg/ml was conducted before the cells were infected with the virus at 37°C. HSV-1 strain gL86 and HSV-2 strain gJ were allowed to infect cell monolayers for 6 hours, after which the cells were washed with PBS twice and 100 μl of soluble substrate o-nitrophenyl-β-d-galactopyranoside (3 mg/ml) was added to the cells along with 0.5% Nonidet P-40 in PBS. Enzymatic activity was measured by a microplate reader (Tecan GENious Pro) at 405/600 nm.

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