Fig. 5.

Intracellular localization of mutant nsp1 proteins. DBT-9 cells grown on glass coverslips were infected with icwt or nsp1 mutant viruses for 7 h, fixed and permeablized with 100% methanol, and incubated with antibodies against nsp1 (red), nsp2 (green), and N (purple). Cells were imaged using a Zeiss LSM 510 confocal microscope at 546 nm (red), 488 nm (green), and 633 nm (purple). Images are single confocal slices obtained using a 40× objective. Co-localization of green and purple for VUSB1 is shown in the merged image as light green pixels. Co-localization of all three colors is shown in the merged images as white pixels. Multi-nucleated cells are a cytopathic effect of MHV replication.