Calcusun

HIV-1 NL4–3 (multiplicity of infection = 0.001) was used to infect MT4 cells in the presence of various concentrations of the test compounds, and the CellTiter-Glo reagent was added in parallel for cytotoxicity studies.⁵ Virus replication was analyzed on day-4 postinfection using a p24 ELISA kit from PerkinElmer, and cytotoxicity was determined using a cell viability kit provided by Promega. The compound concentration that inhibited HIV-1 replication by 50% (EC₅₀) and decreased the cell viability by 50% (IC₅₀) was calculated by using the biostatistic software Calcusun (Biosoft). Gnidimacrin was used as a positive control.

HIV-1 NL4–3 (multiplicity of infection = 0.001) was used to infect MT4 cells in the presence of different concentrations of the test compounds, and cytotoxicity of these compounds to MT4 cells was evaluated by using a cell viability kit provided by Promega.^{8 (link)} Fresh medium containing appropriate concentrations of the compounds was added to the culture 48 h after infection to maintain normal cell growth. Virus replication was analyzed on day-4 postinfection using a p24 ELISA kit from PerkinElmer. The CellTiter-Glo Luminescent Cell Viability Assay is a simple method of determining the viability of the cells in culture based on quantitation of ATP in metabolically active cells. The CellTiter-Glo reagent was added to the MT4 cells that were cultured parallel to the antiviral assays. The biostatistic software Calcusun (Biosoft) was used to calculate the compound concentration that inhibited HIV-1 replication by 50% (EC₅₀) and decreased the cell viability by 50% (IC₅₀). Gnidimacrin was used as a positive control.