FIG. 4.

Summary of the expression of N-terminal deletions of μNS. (a) N-terminal deletions of μNS are indicated as in Fig. 3. The ability of each construct to form intracellular inclusions is indicated as positive (+), negative (−), or aggregated (ag). The immunofluorescence analysis shown on the right was performed as indicated in Fig. 2b on CEFs fixed after 18 h of transfection with plasmids expressing the indicated N-terminal μNS truncations. (b) Ubiquitination analysis of N-terminal deletions. CEFs were fixed after 18 h of transfection with plasmids expressing μNS(140-635) (row 1), μNS(112-635) (row 2), or μNS(381-635) (row 3) as examples of large, aggregated, and small inclusions, respectively. The cells were immunostained with rabbit anti-μNS antibodies (μNS) and mouse anti-conjugated ubiquitin (C-Ubq). The secondary antibodies used were Alexa 488-conjugated goat anti-rabbit IgG (green) and Alexa 594-conjugated goat anti-mouse IgG (red), respectively. In the merged image, colocalization of μNS(112-635) (row 2) and conjugated ubiquitin is indicated by yellow. Nuclei were counterstained with DAPI (blue).