Figure 1.

Confocal microscopy images show differential binding of SNV^R18 to Vero E6 cells in HHB/0.1% HSA buffer in the presence of A. 1.5 mM Ca²⁺ which supports an integrin bent conformation low-affinity state, B. 1 mM Mn²⁺ activates integrins and induces an extended integrin conformation. C. Excess unlabeled SNV used to block SNV^R18. D. Flow cytometry measurements of SNV^R18 binding to Vero E6 cells in the presence Ca²⁺, Mn²⁺, and excess unlabeled SNV. 100,000 Vero E6 /well, were incubated with 1500 SNV^R18/cell at 37°C under mild vortexing for 15 mins. The cells were then washed 3x in the buffer, then either removed from the bottom of the wells by 0.025% Trypsin/0.0526 mM EDTA for flow cytometric measurement or fixed in 2% PFA at 0°C for 30 mins before mounting in Vector Shield containing 1X DAPI for microscopic imaging. E. Equilibrium binding of SNV^R18 titers to 10,000 Tanoue B cells in a 384 well plate. Quadruplicate measurements were performed for each condition to determine the signal to background ratio and Z’ values for each titer. The Z’-score values for each condition were comparable, here Z’ scores for +DMSO are shown. Error bars represent ±SD.