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Fig. 2.
Effects of dexamethasone on RANKL, OPG, and M-CSF mRNA expressions after 12 h of suspension culture. (A–D) Human bone marrow adipocytes were cultured in serum-free DMEM with dexamethasone (10?7 mol/L) for 12 h, and real-time PCR of RANKL, OPG, and M-CSF was performed for RNA extracted from the floating adipocyte layer. Data represent relative expressions against control levels and EPZ005687 expressed as the mean ± SD of measurements. Asterisks indicate significant increases compared with basal levels. RANKL and OPG expressions and the RANKL/OPG ratio are significantly increased (p expression levels for three independent subjects, and are expressed as the mean ± SD of triplicate measurements.
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For the dose-dependent study, real-time PCR was carried out on three independent primary bone marrow adipocyte cultures that were randomly selected from 10 individuals treated with various concentrations of dexamethasone for 12 h. Significant increases in RANKL mRNA expression, OPG mRNA expression, and the RANKL/OPG ratio were observed at 10?7 mol/L (Fig. 2E, F, and G).