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. 2008 Sep 24;82(24):12487–12497. doi: 10.1128/JVI.00877-08

FIG. 6.

FIG. 6.

Determination of the subcellular distribution and colocalization of GABARAP and BDV P protein. (A and B) pEGFP-P or pAsRed2C1-GABARAP was transfected individually into COS-7 cells. The expression and distribution of fusion proteins GFP-P (A) and AsRed2-GABARAP (B) in transfected cells were examined by laser-scanning confocal microscopy. (C and D) pAsRed2C1-GABARAP and pEGFP-P or pEGFP-PΔC1 were cotransfected into COS-7 cells. The colocalization of fusion proteins GFP-P with AsRed2-GABARAP (C) and GFP-PΔC1 with AsRed2-GABARAP (D) in transfected cells was examined by laser-scanning confocal microscopy with appropriate filters. (E) For investigating the colocalization of P and GABARAP in BDV-infected COS-7 cells, cells were infected with BDV and then transfected with pAsRed2C1-GABARAP. Anti-P antibody was used as the first antibody, and anti-rabbit IgG antibody labeled with fluorescein isothiocyanate (FITC) was used as the second antibody. Proteins were examined by laser-scanning confocal microscopy. To show the location of the nuclei, cells were stained with Hoechst 33258. Scale bar, 20 μm.