Figure 7.

ILP1 Represses Ccna2 Expression in Mouse NIH3T3 Cells.

(A) The constructs used for the in vivo transcription assay in mouse NIH3T3 cells. pcDNA-ECFP-40 contains the enhanced cyan fluorescent protein (ECFP) gene and was used as a control, and pcDNA-MusILP1-40 contains the mouse ILP1 cDNA (731 amino acids). The reporter plasmid consists of the Ccna2 promoter region (−177 to +100 bp of the transcriptional start site) fused to the LUC gene. The reference plasmid serves to monitor the transfection efficiency by β-galactosidase (LacZ) expression. CMV pro, Cytomegalovirus promoter; BGH pA, bovine growth hormone polyadenylation site.

(B) In vivo transcription assay in mouse NIH3T3 cells. LUC activity was normalized with β-galactosidase activity. Relative LUC activity: LUC activity of mouse ILP1 relative to enhanced cyan fluorescent protein. Activities were measured 24 and 48 h after transfection. Error bars indicate standard deviation. The experiment was repeated four times.