Fig. 6: CHIR-99021, an activator of Wnt/β-catenin pathway ameliorated the imbalance between osteogenic and adipogenic differentiation in MSCs caused by Mapk7 deletion.

A ARS staining for CHIR-99021 (10 μM) treatment to primary MSCs at passage 4 after OS induction for 14 days, scale bar: 150 μm. B Quantitative of ARS staining, n = 3, ^***p < 0.005. C ORO staining for CHIR-99021 (10 μM) treatment to primary MSCs at passage 4 after OS induction, scale bar: 200 μm. D Quantitative of ORO staining, n = 3, ^***p < 0.005. E Protein expression of Mapk7, osteogenesis-related genes (Runx2), and β-catenin was detected after OS induction of primary MSCs for 3 days with CHIR-99021 (10 μM) treatment. F Application of CHIR-99021 (10 μM) to MSCs at passage 4 after OS induction for 3 days, qPCR was performed to detect the levels of osteogenesis-related genes (Runx2, Alpl), n = 3, ^*p < 0.05; ^**p < 0.01. G Primary MSCs at passage 4 of WT/CKO mice were treated with CHIR-99021 (10 μM) for 3 days after AD induction, and Western blot was performed to detect the expression of Mapk7, adipogenesis-related gene (Fabp4) and active β-catenin, β-actin was used as an internal control. H Adipogenesis of primary MSCs at passage 4 was induced for 3 days along with CHIR-99021 (10 μM) treatment, then qPCR was performed to detect the mRNA levels of lipid formation-related genes (Fabp4, Cebpα), n = 3, ^***p < 0.005.