Figure 4

Translocation of TFPI-2 into nucleus decreased promoter activity of the MMP-2 gene. (A) Representative images show the cellular distribution of TFPI-2 in MDA231 cell lines (left). Scale bars: 10 μm. An average of 100 TFPI-2 overexpression cells was examined and the percentage of the cells with enriched TFPI-2 protein in the nucleus is presented (right). **P < 0.01 as determined by Student’s t-test. (B) Analyses of MMP-2 promoter activity in MDA231 cell clones. Upper: schematic representation of the luciferase reporter driven by the MMP-2 promoter. Arrows indicate the transcription start site for the luciferase gene. Lower: Luciferase reporter driven by the MMP-2-promoter and an internal control Renilla luciferase plasmid were co-transfected into cultured MDA231 cell lines. The firefly luciferase activity was normalized to the Renilla luciferase activity and the relative luciferase activity is represented. The results were the average of a total of three independent experiments each carried out in triplicate. *P < 0.05 as determined by Student’s t-test. (C) Levels of reporter firefly luciferase mRNA were determined by RT-PCR and normalized to Renilla luciferase mRNA for each cell line. Mean values (+SD) from three independent experiments are shown. *P < 0.05, **P < 0.01 as determined by one-way ANOVA followed by Tukey’s multiple comparison tests.