Figure 6.

A. A plot of Green fluorescence of CHO-A24 cells expressing an Upward cAMP Difference Detector in situ (CADDis) sensor, responding to 10 μM forskolin (positive control for cAMP activity) and SNV^R18 alone and mixed with 100 μM Antimycin, 1.5 mM Mn²⁺ and GRGDSP. Error bars represent ±SD for triplicate measurements. B. The plot compares endpoint cyclic AMP signals produced in antimycin treated cells (no wash) and untreated cells (see Fig. S3 for real-time assay). C. SNV^R18 binding to α_IIbβ₃ recapitulates physiologic integrin activation indicated by PAC-1 binding. The graph shows a plot of fold increase over resting cells in PAC-1 staining of CHO-A24 cells after exposure to SNV^R18. 5,000 CHO-A24 cells were suspended in 20 μl media containing 1:20 dilution PAC-1 antibody. Select tubes were treated for 30 min with 10 μM GRDGSP. SNV^R18 samples were premixed with antimycin to block SNV^R18 binding to cells. To induce PAC-1 staining SNV (5,000 particles/cell) were added to the tubes and incubated with virus for 5 min. Cell activation was terminated by placing tubes in ice-cold water. PAC1 was allowed to equilibrate for 30 min. Cells were washed once and read on a flow cytometer. Error bars represent standard error of 3 triplicate measurements. Statistical significance of the difference between the vehicle (Veh) and compound-treated samples was determined by Dunnett multiple comparison t-tests.