Fig. 3

GW4064 treatment inhibited cell proliferation and induced apoptosis and cell cycle arrest in MG-63 cells. We further study the effects of farnesoid X receptor (FXR) regulation on the development of MG-63 cells and the role of microRNA-23b-3p during the progress. a, b In order to determine the effects of FXR regulation on osteosarcoma (OS) cell proliferation, clone formation assay of MG-63 cells were performed under the single and combined effects of miR-23b-3p inhibition and GW4064 treatment. c, d The changes of apoptosis rate were analyzed by flow cytometry. e, f The cell cycle distribution of MG-63 cells was determined by flow cytometry. g, h The protein levels of several apoptotic genes (B-cell lymphoma-2 (Bcl-2), Bax, and Cleaved Caspase-3 (C caspase-3)) were analyzed by Western blot to further assess the effects of FXR regulation on OS cell apoptosis. Each value represents mean ± SEM (n = 3). GAPDH served as an internal control. *p < 0.05, **p < 0.01 vs. IC group; ^##p < 0.01 vs. IC + GW4064 group; ^^p < 0.01 vs. I group