Effect of inducible aNotch1 in ST2 stromal cells on their ability to support osteoclastogenesis.
After culturing for 72 hours at the indicated concentration of Tc, the expression of aNotch in ST2NIC stromal cells was assessed by measuring the fluorescence intensity of GFP (A). (B) Freshly prepared BM cells were cultured for 6 days on ST2NIC with Tc (100 ng/mL) or the same volume of EtOH in the presence of 1.25(OH)2D3 (10−8 M) and DEX (10−7 M). After 6 days of culturing, TRAP+multinucleated osteoclasts were counted. Bars indicate means ± SD of triplicate cultures. *Indicates significantly different from the corresponding cultures with EtOH (P < .001). (C) After culturing for 4 days with Tc (100 ng/mL) or EtOH in the presence of 1,25(OH)2D3 and DEX (D/D), Northern hybridization analysis of Csfm (M-CSF), Tnfsf11(RANKL), and Tnfrsf11b (OPG) gene expression in ST2NIC was performed. Gapd gene expression is shown as a loading control in the bottom blot.
