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. 2009 Oct 12;25(4):819–829. doi: 10.1359/jbmr.091019

Fig. 6.

Fig. 6

PTH induction of COX-2 expression and promoter activity depends on calcium-calcineurin signaling. MC-4 cells stably transfected with Pluc371 were treated with vehicle, PTH (10 nM), or forskolin (FSK, 1 µM) for 1 hour (mRNA) or 3 hours (luciferase activity). BAPTA-AM (BAPTA, 15 µM), FK506 (10 µg/mL), cyclosporin A (CsA, 10 µg/mL), or VIVIT (1 µM) were added 1 hour before PTH or forskolin. Vehicle (CON) for the inhibitors, DMSO, also was added 1 hour before PTH or forskolin. Luciferase activity was normalized to total protein and used to calculate the fold induction as the ratio of treatment to control. (A) Effects of the calcium chelator BAPTA-AM and the calcineurin inhibitors FK506 and CsA on PTH-induced COX-2 mRNA expression measured by real-time RT-PCR. (B) Effect of BAPTA-AM and the calcineurin inhibitors FK506, CsA, and VIVIT on the PTH induction of luciferase activity. (C) Effect of BAPTA-AM, FK506, and CsA on the forskolin induction of luciferase activity. Bars are means ± SEM for n = 3 wells of cells. aSignificant effect of agonist, p < .01. bSignificant effect of inhibitor, p < .01.