Figure 3

Pimecrolimus exhibits antiviral activity against DENV2 infection, (a) Viability of Vero cells treated with the indicated concentrations of pimecrolimus or ribavirin without virus for 4 dpi. (b) Vero cells infected with 0.1 MOI of DENV2 and treated with the indicated concentrations of pimecrolimus and ribavirin for 4 dpi. (c) Representative images of the crystal violet staining for visually comparing the viability of control and DENV2-infected Vero cells across the different compound concentrations used. (d) BHK-21 cells treated with the indicated concentrations of pimecrolimus and ribavirin without virus for 2 dpi. (e) BHK-21 cells infected with 0.1 MOI of DENV2 and treated with the indicated concentrations of pimecrolimus and ribavirin for 2 dpi. (f) Representative images of the crystal violet staining for visually comparing the viability of control and DENV2-infected BHK-21 cells across the different compound concentrations used. (g) Plaque reduction assay in DENV2-infected BHK-21 cells treated with the indicated concentrations of pimecrolimus and ribavirin. After 4 days, cells were stained with 0.2% crystal violet, and representative images of cell viability results are shown. (h) Comparison of the relative number of plaques formed in BHK-21 cells across the different pimecrolimus or ribavirin treatment concentrations used. (i) Plaque reduction assay in DENV2-infected Vero E6 cells treated with the indicated concentrations of pimecrolimus and ribavirin. After 7 days, cells were stained with 0.2% crystal violet, and representative images of cell viability results are shown. (j) Comparison of the relative number of plaques formed in Vero E6 cells across the different pimecrolimus or ribavirin treatment concentrations used. (k) Plaque reduction assay on DENV2-infected Huh-7 cells treated with the indicated concentrations of pimecrolimus and ribavirin. After 7 days, cells were stained with 0.2% crystal violet. Representative images of cell viability results are shown. (l) Comparison of the relative number of plaques formed in Huh-7 cells treated with different pimecrolimus and ribavirin concentrations. 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 group).