FIG 5.
Subcellular distribution of the mutant Z proteins in the presence or absence of an active vRNP. 293T cells seeded on coverslips in a 24-well plate (1 × 105 cells/well) and cultured overnight were transfected with 25 ng of plasmid expressing WT or the indicated mutant Z proteins together with (B) or without (A) plasmids required for intracellular reconstruction of an active vRNP (vRNP) (pT7-MG/CAT, pC-T7pol, pC-L, and pC-NP). Forty-eight hours later, transfected cells were fixed and Strep-tagged Z proteins and HA-tagged NP expression detected by IF using a mouse monoclonal antibody to Strep and a rabbit polyclonal antibody to HA, respectively. Nuclei were detected by DAPI staining. Stained cells were analyzed with a confocal microscope.