Figure 5. Y1R antagonist treatment reversed NPY-mediated suppression of osteogenesis and cAMP/PKA/CREB pathway in BMSCs. (A) Flow cytometric analysis of BMSC surface markers. (B) The multidifferentiation potential of BMSCs in vitro (Alizarin red S staining for osteogenic differentiation, Alcian blue staining for chondrogenic differentiation, and oil Red O staining for adipogenic differentiation). (C) Western blotting results of OPG, RANKL, RUNX2, OCN, p-PKAs and p-CREB expression in BMSCs after 3 days of incubation in osteogenic medium with NPY or NPY+BIBO3304. (D, E) The cAMP concentrations and ALP activity were measured on days 3 of incubation in osteogenic medium with NPY or NPY+BIBO3304. The data are expressed as the means ± SD. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001 vs. CON, and #P<0.05; ##P<0.01; ####P<0.0001 vs. NPY by one-way ANOVA and Tukey’s post hoc test.