Figure 6
From: Sclerostin expression in trabecular bone is downregulated by osteoclasts
Expression of LIF in RANKL-stimulated BMMs and osteoclasts. (A) BMMs were cultured in the presence or absence of GST-RANKL (100 ng/ml), GM-CSF (10 ng/ml), IL-4 (10 ng/ml), or IFN-γ (20 ng/ml) with M-CSF (50 ng/ml) for 24 h. Analysis of the expression of Lif and Ctsk mRNAs in the cultured BMMs using real-time PCR (n = 3). (B) BMMs were cultured in the presence or absence of GST-RANKL (100 ng/ml) with M-CSF (50 ng/ml) for 3 days. Osteoclasts (OCLs) appeared on day 3. In addition, the OCLs were further cultured in the presence of varying concentrations of GST-RANKL (0, 1, 5, 25, or 100 ng/ml) with M-CSF (50 ng/ml) for 24 h. For inhibitor experiments, the OCLs were further cultured in the presence or absence of inhibitors of JNK, p38 MAPK, ERK, or NF-κB pathways with GST-RANKL (100 ng/ml) and M-CSF (50 ng/ml) for 24 h. (C) Analysis of the expression of Lif, Acp5, Ctsk, and Mmp9 mRNAs in the cultured OCLs in the presence of varying concentrations of GST-RANKL (0, 1, 5, 25, or 100 ng/ml) with M-CSF using real-time PCR (n = 4). (D) Analysis of the expression of Lif mRNA in the cultured OCLs in the presence or absence of inhibitors of JNK (10 μM), p38 MAPK (10 μM), ERK (20 μM), or NF-κB (5 μM) pathways with GST-RANKL (100 ng/ml) and M-CSF using real-time PCR (n = 4). *p < 0.05, **p < 0.01.
