LNShh cells do not upregulate Runx2 expression in MC3T3 cells. (A) MC3T3 cells were grown in mixed cultures with either LNCaP or LNShh cells. Expression of Runx2, Ocn, and Opn in MC3T3 cells cultured with LNShh cells (filled bars) relative to those cultured with control LNCaP cells (open bars) were compared. (B) MC3T3 cells were cultured alone for 24 h without (-) or with (+) 1 μg/ml Shh-N or 50 μg/ml AA as indicated. Levels of expression of Runx2, Gli1 and Ptc1 among groups were compared. All values are means ± SD from 2–3 independent experiments. *, P < 0.05.