Figure 6.

NADH levels determine Runx2 target gene expression. (A) The NAD⁺/NADH levels in AOB cells were significantly increased. Four HOB cell samples (1-4) and four AOB cell samples (1-4) were subjected to measure the NAD⁺/NADH levels. **P<0.001. (B) The CoCl₂ treatment dramatically decreased the NAD⁺/NADH levels. The HOB1 and AOB cells were treated with CoCl₂ or CoCl₂+HQNO, and then, the NAD⁺/NADH levels were measured. **P<0.001. (C) Treatments with CoCl₂ or CoCl₂+HQNO cannot affect the CtBP2 mRNA level. The mRNAs from cells used in (B) were subjected to qRT-PCR analysis to determine the CtBP2 levels. **P<0.001. (D) The CoCl₂ treatment significantly increased Runx2 acetylation. Cells used in (B) were subjected to western blot to examine the CtBP2, p300, Runx2 and acetylated-Runx2 levels. GAPDH was used as a control. (E) The CoCl₂ treatment significantly increased the expressions of Runx2 target genes. The mRNAs used in (C) were subjected to qRT-PCR analysis to determine the expressions of Runx2 target genes including OSC, ALPL, SPP1, COL1A1, IBSP and MMP13. CUL4A was used as a control. **P<0.001.