Fig. 5.

Dlx3 occupies the human MMP-9 gene promoter endogenously. A: analyses of the human MMP-9 gene promoter revealed 3 near consensus Dlx3 binding sites containing the core TAATT motif within a 2-kb portion of the 5′-flanking region specifically located at −1,812, −1,268 and −889 relative to the transcriptional start site of this gene. B: chromatin immunoprecipitation (ChIP) studies were carried out in JEG3 cells expressing Dlx3 endogenously. Several separate targets were assayed by PCR following ChIP. As a positive control, the junctional regulatory element from the human α-subunit gene promoter (hα JRE) was used; a target ∼2 kb downstream of the JRE within the α-subunit gene was used as a negative control (hα Distal); the putative Dlx3 binding at position −889, −1,268, and −1,812 were selected within the human MMP-9 gene promoter. For each condition, a ChIP reaction was carried out with protein A beads alone or with an IgG control antibody to serve as controls. This ChIP study was carried out on at least 3 separate occasions, each in triplicate, with similar results. *P ≤ 0.05, increased MMP9 promoter occupancy with Dlx3 overexpression.