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. 2012 Sep 4;122(10):3490–3503. doi: 10.1172/JCI64906

Figure 2. Serotonin suppresses osteoblast proliferation through FOXO1.

Figure 2

(A) SOD2 activity in osteoblasts treated with serotonin (5-HT; 100 μM) for 6 hours (n = 3). *P < 0.05 versus vehicle. (B) Calvaria cells cotransfected with Lrp5 and FOXO-Luc plasmids and treated with vehicle or 100 μM serotonin for 24 hours (n = 3). EV, empty vector. *P < 0.05 versus empty vector, #P < 0.05 versus FOXO-Luc vehicle. (C) Real-time PCR analysis of the expression of Ccnd1 and Ccnd2 in calvaria cells treated with serotonin (100 μM) for 6 hours (n = 3). *P < 0.05 versus vehicle. (D) Calvaria cells from WT and Foxo1ob–/– mice were treated with serotonin (100 μM) for 48 hours, and proliferation was assessed by Cell Titer One (left) or BrdU incorporation (right) (n = 3). *P < 0.05 versus vehicle. (E and F) Calvaria cells from WT and Foxo1ob–/– mice were treated with serotonin (100 μM) for 48 hours. Gene expression of (E) Ccnd1 and (F) Ccnd2 (n = 3). *P < 0.05 versus vehicle. (G) Western blot analysis showing phosphorylation of PKA in WT and Foxo1-deficient osteoblasts treated with serotonin (100 μM) for 30 minutes. (H) Serum serotonin levels (n = 4 mice/group). *P < 0.05 versus WT, #P < 0.05 versus Lrp5+/–Foxo1ob–/– group.