Figure 2: 5-HT6R inhibits osteoblast differentiation via the BMP-2 signaling pathway but not Gs-protein signaling. | Scientific Reports

Figure 2: 5-HT6R inhibits osteoblast differentiation via the BMP-2 signaling pathway but not Gs-protein signaling.

From: Peripheral serotonin-mediated system suppresses bone development and regeneration via serotonin 6 G-protein-coupled receptor

Figure 2

(A,B) Primary calvarial osteoblasts from 1-day-old ICR mice were cultured with OS in the presence of 10 μM ST1936 for 0, 1, 3, and 7 days. Phospho-Smad1/5/8 and Smad1/5/8 (A), and Runx2 (B) were assessed by Western blot analysis. (C–F) Cells were cultured in OS with rh-BMP2 in the presence of ST1936 for 3 (C,D) and 10 days (E,F). ALP activity was measured via ALP staining (C) and ALP activity (D). Mineralized nodule formation was assessed by Alizarin Red staining (E), and then optical density was measured at 590 nm. (F). (G–J) Cells were pretreated with H89 (1 μM) or U0126 (1 μM) for 1 h, and then cultured in OS with ST1936 for 5 days (G,H) and 14 days (I,J). ALP activity was measured via ALP staining (G) and ALP activity (H). Mineralized nodule formation was assessed by Alizarin red staining (I) and the stains were eluted and measured at 590 nm. (J). Data shown represent the means ± SEM. *p < 0.05, vs. control. #p < 0.05, vs. OS. &, p < 0.05, vs. ST1936.

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