Figure 8.

The effect of the DENV2C mutations on capsid-RNA interaction was evaluated. (A) Interactions of BSA (negative control), DENV2C WT and L50S, L54S, L81N and I88N mutants with a nonspecific RNA were measured by fluorescence polarization. Reactions were prepared containing 50 nM RNA-FAM, RNase inhibitor (diluted 1:5000), and protein concentrations ranging 0 to 10 µM in 50 mM sodium phosphate/20 mM NaCl, at a reaction volume of 100 µL. The data represent the means of three individual experiments and were fitted by non-linear regression analysis to a one-phase association equation using the least squares method available in GraphPad Prism 7 software. (B) The anisotropy value for each protein concentration was also converted into a binding value, which was plotted against protein concentration.