FIG. 9.

1,25(OH)₂D₃ induces histone acetylation in the upstream regions of the mRankL gene. ST2 cells were treated with 1,25(OH)₂D₃ (10⁻⁷ M) for periods of up to 6 h and then subjected to ChIP analysis using antibodies to VDR, tetra-acetylated histone 4, or IgG (αVDR, αAcH4, and αIgG, respectively). Precipitated DNA was isolated and evaluated by PCR using Cyp24a1 primers (see Materials and Methods) or RankL primers whose locations are depicted in Fig. 3 (upper panel) and whose sequences are delineated in Table 1. Amplifications were carried out as in Fig. 3. Similar results were obtained in at least four separate experiments.