Figure 2

Pimecrolimus inhibits DENV2 replication at an early stage of infection. (a) Experimental scheme of the time-of-addition assay. (b) Vero cells infected with 0.1 MOI of DENV2. Pimecrolimus at 20 μΜ was added at the indicated time points, and genome expression was analyzed 12 h post-infection using real-time PCR. (c) Binding inhibition assay and (d) internalization inhibition assay of 0.1 MOI DENV2-infected Vero cells treated with the indicated concentrations of pimecrolimus at 96 hpi using cell viability assay. (e) Binding inhibition assay and (f) internalization inhibition assay of 0.1 MOI DENV2-infected Vero cells treated with the indicated concentrations of pimecrolimus at different time points via real-time PCR for DENV2 genome copy number. Data are expressed as mean ± SEM (n = 3 independent biological experiments, each conducted with triplicate technical repeats; ns, not significant; ***p < 0.001; comparisons were made against the DENV2-infected and non-infected control groups.