Figure 3

V38E mutant fails to induce bystander apoptosis in vivo. Spleens isolated 8 weeks post infection were fixed in formalin and sectioned. Paraffin embedded sections were stained with anti-p24 RD-1 antibody (red), active caspase 3 antibody (green) and the nuclear stain, DAPI (blue). Individual channels and merge images for WT (A) and V38E mutant (B) infected mice spleens are shown. Enlarged images (right most) from A and B show the presence of apoptotic cells in close proximity to infected cells in WT, but not V38E infected mice. (C) Automated fluorescence quantitation of total (DAPI), apoptotic (Caspase) and infected cells (p24) from at least 6 images from 2 different slides from each mice was performed using NIS elements image analysis software (Nikon). Each symbol represents an individual mouse and the horizontal lines represent the mean.