Figure 3.

Mechanistic profiling of GRP-156784. A, time-of-addition variation studies against the RSV target. Compound GRP-156784 (red), RSV entry inhibitor BMS-433771 (blue), or viral polymerase inhibitor JMN3–003 (maroon) were added to equally infected cells at the indicated time points and concentrations. Relative virus replication in all cultures was determined 33 h after infection. B, dose–response minigenome reporter assay inhibition by GRP-156784. The compound was tested against RSV–, IAV-WSN–, and MeV–derived minigenomes. C, RT-qPCR quantitation of relative RSV N protein–encoding mRNA levels present in RSV-infected cells after incubation in the presence of GRP-156784, RSV ribonucleoside analog inhibitor ALIOS-8176, or vehicle volume equivalents. Statistical analysis through one-way analysis of variance with Dunnett's multiple-comparison post hoc test. Symbols in A–C represent individual biological repeats (n = 3 each); error bars, S.D. Previous characterized reference compounds BMS-433771 and JMN3–003 were analyzed in A in two biological repeats; error bars, range. In C, each of the three biological repeats was determined in technical duplicates.