Figure 1
Nlz1 represses transcription in various cell lines. A. Western blot demonstrating that GAL4DBD (lane 2) and the Nlz1-GAL4DBD fusion protein (lane 3) are well expressed. No TF = untransfected control. B. Schematic outline of constructs used. The reporter construct (Top) contains three UAS elements (GAL4 binding sites) upstream of the SV40 promoter and the Luciferase reporter gene. The Nlz1-GAL4DBD construct (bottom) was generated by fusing the GAL4DBD in frame to the C-terminus of full-length zebrafish Nlz1. SP = Sp domain; Btd = buttonhead domain; ZF = C2H2 zinc finger domain. C-E. HeLa cell reporter assays were carried out as described in the Methods section. Luciferase activity was normalized to β-galactosidase activity and the data are presented as fold activation relative to control vector. C. The Gal4DBD domain is a weak activator. D. Nlz1 represses transcription from the UAS-SV40:Luciferase reporter in HeLa cells. E. Nlz1 represses transcription in ZF4, SN17 and NIH3T3 cells.