Fig. 2

An HFD induces BMSC senescence and suppresses BMSC osteogenesis. a Representative images following SA-β-gal, EdU, and p21 immunofluorescence staining and alizarin red staining of BMSCs from HFD-fed mice and ND-fed mice (scale bars = 50 μm). b Intracellular ROS levels in BMSCs were detected by DHE staining and flow cytometry. c DNA damage markers in BMSCs from HFD-fed mice and ND-fed mice were detected via immunofluorescence staining of γH2A.X (scale bars = 50 μm). d CCK8 results showing the proliferative activity of BMSCs from HFD-fed mice and ND-fed mice. e Quantification of SA-β-gal, EdU, P21, and ARS staining in the BMSCs from ND- and HFD-fed mice. f Statistical analysis of the DHE staining results. g Quantification of γH2A.X-positive cell ratios in the BMSCs from ND- and HFD-fed mice. h The expression of the cyclin-dependent kinase inhibitors p21, p53, and p16 in BMSCs from HFD-fed mice and ND-fed mice was detected by RT‒qPCR. i Osteogenesis-related gene expression was quantified by RT‒qPCR after osteogenic induction. The data were shown as the mean ± SEM. N = 3, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1