DNA demethylation at the Sp7 promoter accompanies long-term expression of the Sp7 gene. (A) BMP2 treatment results in transient and stable Sp7 gene expression in MB and UD mesenchymal cells, respectively. Sp7 mRNA levels were determined by RT-qPCR as explained in the legend to Fig. 1 and are presented as values relative to the values for the corresponding untreated cells. (B) Treatment of MB cells with BMP2 for 48 h does not induce DNA demethylation at the Sp7 promoter. (C) Histone H3 enrichment at the Sp7 promoter is not reduced in MB cells treated with BMP2 (48 h). (D) MB cells incubated with BMP2 exhibit increased enrichment of H3Ac but not of H3K4me3 at the Sp7 promoter. (E) Enrichment of RNAPII, Dnmt1, and Tet2 (but not Tet1) at the Sp7 gene promoter in MB cells treated with BMP2 (48 h). Data represent means ± standard errors of the means (n ≥ 3). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001 (as determined by Student's t test).