FIG 3.

DENSpm inhibits RNA virus replication. (A and B) Time course of infection of Vero-E6 cells treated with 10 μM DENSpm for 16 h prior to infection with CVB3 (A) or VSV (B) at an MOI of 0.1 or 0.01, respectively. Titers were determined at the times indicated. (C to E) Vero-E6 cells were pretreated with escalating doses of DENSpm for 16 h prior to infection with CVB3 at an MOI of 0.1 (C), VSV at an MOI of 0.01 (D), or CHIKV at an MOI of 0.1 (E). Titers were determined at 24 hpi (40 h posttreatment). (F) Cell counts (open circles, left axis) and viability (closed circles, right axis) were measured for uninfected Vero-E6 cells treated with escalating doses of DENSpm for 40 h. Dashed lines indicate levels corresponding to the results for untreated cells. (G) Western blot analysis of SAT1 and GAPDH (top two panels) and thin-layer chromatographic analysis of polyamines (bottom panel; polyamines labeled for spermidine [Spd], spermine [Spm], and putrescine [Put]) were performed on uninfected samples run, measured at 40 h posttreatment. Statistical significance was determined using Student's t test (n ≥ 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001. Error bars represent 1 standard error of the mean.