Fig. 6: O-GlcNAcylation of FOXO3-Ser296, which is regulated by GLUL, promotes the osteogenic differentiation of BMSCs.

A Volcano plots of BMSCs from Glul^Prx1 (n = 3) and Glul^fl/fl (n = 3) mice subjected to 4D DIA O-GlcNAcylation profiling analysis. B Mass spectrometry image of the O-GlcNAcylation modification of the FOXO3 Ser296 residue. C The results of the co-IP experiment on the effect of Glul knockout on the level of O-GlcNAcylation modification of FOXO3 in BMSCs. D Representative images of target protein decay at different time points in BMSCs from Glul^Prx1 and Glul^fl/fl mice. E Representative images of target protein attenuation at different time points were obtained by transfecting wild-type or S296V-mutant FOXO3 plasmids into Foxo3-KO BMSCs. F Representative images of ARS and ALP staining of Foxo3-KO BMSCs after transfection with wild-type or S296V-mutant FOXO3 plasmids. G Quantitative analysis of ARS and ALP staining. H Representative images of FOXO3-Ser297 O-GlcNAcylation levels as determined by Western blotting analysis of BMSCs from patients in the SOP and control groups. I Western blotting analysis of the expression of osteogenesis-related genes in Foxo3-KO BMSCs after transfection with wild-type or S296V-mutant FOXO3 plasmids. J Representative images of ROS detection. **P < 0.01 vs. other groups; *P < 0.05 vs. other groups.