Lithium and Copper Induce the Osteogenesis-Angiogenesis Coupling of Bone Marrow Mesenchymal Stem Cells via Crosstalk between Canonical Wnt and HIF-1α Signaling Pathways

Figure 6

Lithium and copper costimulation can promote osteogenesis and angiogenesis via crosstalking between the canonical Wnt and HIF-1α signaling pathways. (a) The BMSCs were exposed to 25 μM copper, 500 μM lithium, and costimulation. The Alizarin Red staining was performed to investigate the calcium nodules after four weeks of stimulation. The immunofluorescence (IF) staining was used to detect the VEGF expression in the BMSCs after 7 days of coculture. (b, c) The osteogenesis (ALP, osterix) and angiogenesis (VEGF) genes were detected by RT-qPCR and western blot in the BMSCs under 25 μM Cu²⁺, 500 μM Li⁺, and costimulation intervention for 7 days. (d, e) ChIP analysis for HIF-1α, β-catenin, osterix, and VEGF promoter binding site following stimulation with 25 μM copper and 500 μM lithium or costimulation for 7 days. holes per group. Data shown are of a single representative experiment with an experiment repeated two times with three technical replicates in each experiment. The results are depicted as the . , , and .

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