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5 cells/mL) in RPMI 1640 medium containing 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin were infected with HIV-1 strain IIIb at a multiplicity of 0.01 and were distributed into 96-well microtiter plates. Test compounds were added to the wells, and cultures were incubated at 37 °C for 5 days. Cell viability was determined by the MTT assay, which measures living cells on the basis of mitochondrial dehydrogenase activity. The cells were incubated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution (7.5 mg/mL). The MTT formazan crystals were dissolved in acidic 2-propanol containing 4% Triton-X, and the absorbance was measured at 595 nm.
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