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. Author manuscript; available in PMC: 2011 Jul 19.
Published in final edited form as: Vaccine. 2010 Jun 4;28(32):5280–5287. doi: 10.1016/j.vaccine.2010.05.054

Figure 1. Schematic representation of the plasmids used in this study and in-vitro expression of these plasmids in HEK293 cells.

Figure 1

A The binary plasmid system contains two plasmid vectors. CMV-Gal4 is the activator plasmid, in which, Gal4 was cloned into a pCMVi mammalian expression vector containing CMV promoter for initiation and SV40 polyadenylation site for termination of transcription. The effector plasmids include UAS-Aβ42 monomer or Aβ42 trimer, which contain four UAS sequences for Gal4 binding and transcription initiation and the SV40 polyadenylation sequence for termination. Aβ42 were cloned between adenovirus E3 leader signal and endosomal targeting (ET) sequence. B For comparison with the binary plasmid system, Aβ42 trimer was cloned directly downstream of a CMV promoter. All plasmids contain an intron sequence to further increase transcription. C Western Blot for detection of Aβ42 using the monoclonal antibody 6E10. No band is detected in cells transfected with a control plasmid (Lane C1). After transfection with the respective plasmids in HEK293 cells positive bands of about 30 kDa for pUAS-Aβ42 monomer (Lane C2), 40 kDa for pUAS-Aβ42 trimer (Lane C3) and 40 kDa for CMVi-Aβ42 trimer (Lane C4) confirmed expression of the constructs. D Gal4 expression in CMV-Gal4 transfected HEK293 cells was demonstrated by Gal4 antibody binding to a band of 80 kDa (Lane D2). This band was not detected in cells transfected with control plasmid (Lane D1).