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. Author manuscript; available in PMC: 2010 Apr 1.
Published in final edited form as: Exp Cell Res. 2009 Jan 18;315(6):996–1007. doi: 10.1016/j.yexcr.2009.01.003

Fig. 6.

Fig. 6

The vimentin rod domain cannot substitute for the lamin A rod for repression in mammalian cells. (A) Repression of luciferase reporter by Gal4DBD- fusion proteins of intermediate filament chimeras. Chimeras were made by interchanging the domains between lamin A and vimentin. The graph presents the fold repression of the fusion proteins over GAL4DBD empty vector alone. The data shown are representative of at least three independent experiments. (B) Western blot analysis was performed to verify the expression of fusion proteins. Protein samples were separated by using SDS-4-20% PAGE and detected by immunoblotting with mouse anti-Gal4DBD. The relative protein levels were determined by counting pixel levels using NIH ImageJ and normalizing to beta-actin loading control. (C) Indirect immunofluorescence of 293 T cells expressing Gal4DBD-fusion proteins. Gal4DBD-fusion proteins (red) were detected with antibodies against the Gal4-DNA binding domain. Phalloidin (green) labels actin filaments indicating the cytosol. DAPI (blue) labels DNA indicating the nucleus. Images were acquired using equal exposure times and image processing.