fig7

Figure 7. Analysis of mRNA expression of MMP-9 and the effect of sequence-specific interaction of CD44-ICD/RUNX2 on the promoter region of the MMP-9 gene. A: real-time PCR analysis of MMP-9 expression was done in PC3 cells expressing indicated CD44-ICD deletion constructs. GAPDH was used as a loading control for real-time PCR analysis; B: ChIP assay (top): ChIP assay was performed in cells expressing indicated constructs for MMP-9 promoter. ChIP assay showed an increase in signal in PC3 cells expressing FL, D1, D2, and D3. Signal is considerably reduced in D4 and D5. Immunoblotting analysis with an antibody to MMP9 is shown (middle panel in B). The expression levels of MMP-9 protein corresponded to the observations shown in ChIP assay. Immunoblotting with a sCD44 antibody was used as loading control (B; bottom panel). *P < 0.05 vs. D4 and D5. CD44: Cluster of differentiation 44; ICD: intracellular domain