Fig 6.

Mutations in the RNA-binding domain of the VEEV capsid protein affect the stability of nucleocapsids. The schematic representation of these experiments is shown as part of the diagram in Fig. 5A. (A) Western blotting of viral particles treated under different conditions. (B) Analysis of particle protein content after incubation at 56°C for 1 h and pelleting by ultracentrifugation. (C) Analysis of glycoprotein/capsid ratio in viral particles before and after incubation for 1 h at 56°C. The concentration of proteins on a quantitative Western blot was determined in fluorescence units. (D) Comparison of infectious titers of samples before and after 1 h of incubation at 56°C. (E) Analysis of nucleocapsid stability after Triton X-100 treatment (see Materials and Methods for details). Panel A shows a representative image from one of three highly reproducible experiments; the quantitative data from all three experiments were used for calculation of means and standard deviations for the other panels.