FIGURE 3.

Target of teicoplanin is located within the host cells. A, p24-normalized (50 ng) HIV-luc/Zaire EBOV-GP (2014) pseudotyped viruses were incubated with 50 μm teicoplanin in a 96-well plate at 37 °C for 12 h. Then the compound virus mixtures were transferred into a Microcon 30-kDa centrifugal filter device (Millipore) and centrifuged (7000 × g) at 4 °C for 15 min. Subsequently, fresh medium was added twice onto the filter device to wash the compound virus mixtures. Then 0.5 ml of DMEM was used to suspend the compound virus mixtures, and the reversed filter device was centrifuged (500 × g) at 4 °C for 5 min. The solution was collected and used to infect HEK293T cells after HIV-1 p24 normalization. After 48 h, the intracellular luciferase activity was measured. B, HEK293T cells were incubated with teicoplanin at various concentrations for 12 h. The cells were then infected with HIV-luc/Zaire EBOV-GP (2014) pseudotyped viruses. After washing and incubation with fresh medium for 48 h, the intracellular luciferase activity was measured. The assay during treatment was the same as the antiviral entry assay. The results are representative of at least three independent experiments. The bars show the mean values ± S.D. (error bars). The p value was determined by a Student's t test. n.s., not significant; **, p < 0.01; ***, p < 0.001.