Fig. 7

Azoramide inhibited β-catenin nuclear translocation. a Azoramide (Azo) treatment inhibited β-catenin nuclear localization. C3H10T1/2 cells were pretreated with osteogenic differentiation medium for 12 h followed by azoramide treatment for 6 h and cells were then fixed for immunofluorescence with anti-β-catenin antibody. Scale bar = 20 μm. b Western blot of phosphorylated β-catenin in C3H10T1/2 cells stimulated by 15 μM azoramide at the indicated times. c Western blot of TCF7L2 mRNA in C3H10T1/2 cells subjected to different treatments. Cells were cultured in osteogenic differentiation medium. d Western blot of Runt-related transcription factor 2 (Runx2) and peroxisome proliferator-activated receptor gamma (PPARγ) proteins in C3H10T1/2 cells treated with azoramide and Wnt agonist 1, a small-molecule agonist of the Wnt/β-catenin signaling pathway. e Alizarin red S staining for mineral deposition and Oil red O staining for lipid production of cells subjected to different treatments. f Western blot of phosphorylated β-catenin in C3H10T1/2 cells subjected to different treatments. g Model representing the azoramide role in regulatory effects on C3H10T1/2 cell differentiation. *p < 0.05, **p < 0.01, versus control (Con); ^#p < 0.05, versus Azo. glp-1r glucagon-like peptide-1 receptor, si small interfering