Unveiling the role of hypoxia-inducible factor 2alpha in osteoporosis: Implications for bone health

Figure 6 Treatment with the hypoxia-inducible factor 2alpha agonist roxadustat increases osteogenesis and decreases adipogenesis. A: Representative images of the fibroblast colony-forming unit (CFU-F) assay with Prx1-Cre;Hif-2α^fl/fl and Hif-2α^fl/fl mice are shown; B: Quantification of CFU-Fs formed by bone mesenchymal stem cells (BMSCs) from Prx1-Cre;Hif-2α^fl/fl and Hif-2α^fl/fl mice are shown; C and D: Hif-2α^fl/fl mouse BMSCs were cultured with or without roxadustat. In the adipogenic differentiation assay, Oil red O staining was performed after seven days (D); quantification of Oil Red O staining in BMSCs after induction of adipogenic differentiation (C); E-M: Relative mRNA levels of the adipogenesis-related genes Lpl (E), Adiponectin (F) and Cebpa (G) in BMSCs after 7 d of adipogenic differentiation. In the osteogenic differentiation assay, Alizarin red staining was performed after 14 d to quantify osteoblast mineralization (I). Relative mRNA levels of the osteogenesis-related genes OCN (H), Runx2 (J) and Col1α1 (K) in BMSCs after 14 d of osteogenic differentiation. Adipogenic differentiation was induced in BMSCs by treatment with roxadustat and MHY1485 for 7 d. The levels of the mechanistic target of rapamycin (mTOR) and PI3K/AKT signaling pathway proteins were measured via western blotting in the Ctrl, roxadustat and Roxadustat + MHY1485 groups (M). Quantification of the relative levels of the mTOR, PI3K and AKT proteins (L). The phosphorylated protein levels were normalized to the corresponding total protein levels. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. Roxa: Roxadustat; MHY: MHY1485; HIF-2α: Hypoxia-inducible factor 2alpha; mTOR: Mechanistic target of rapamycin.