Figure 4

Verification of Slc9a3r1 and Sdh1 as HDI-inducible genes. MC3T3 (A and C) or primary calvarial osteoblasts (B and D) were cultured in osteogenic medium containing the indicated HDI or DMSO. mRNAs were isolated at various times over a 6 or 18 hour period and subjected to quantitative real-time PCR with primers for Slc9a3r1 (A and B) or Sdh1 (C and D). Values are relative to those obtained from DMSO-treated samples at each time point and represent the mean of three samples. For (A-D), * denotes a statistically significant change of p < 0.01 and ** denotes p < 0.05 by one-way ANOVA of the HDI-treated sample versus the DMSO-treated sample at that time point. (E) C2C12 cells were cultured in osteogenic medium and 20 nM TSA for the indicated times. Slc9a3r1 protein levels were determined by immunoblotting.