Fig. 5.

DOR deficiency decreases the gene expression of osteoblast-specific markers. Scramble control and DOR shRNA MC3T3-E1 cells were incubated in the presence or absence of T₄ (100 ng/ml) for 24 (osteocalcin) or 72 h (ALP and Runx2). All treatments contained ascorbic acid (100 μg/ml) and β-glycerophosphate (10 mM). Expression of osteocalcin (A), ALP (B), and Runx2 (C) was determined by real-time RT-PCR. Values are presented as fold change ± SE (n = 4). ^AP < 0.05 vs. vehicle control; ^BP < 0.05 vs. scramble control. D: DOR is localized to the nucleus. DOR expression was evaluated by Western blot in MC3T3-E1 cells incubated in the presence or absence of T₃ (10 ng/ml) or T₄ (100 ng/ml) for 1 h. Cell lysates from cytosolic and nuclear fractions were analyzed, and histone H3, β-actin, and β-tubulin served as internal controls. Data are representative of 4 independent experiments (n = 4).