Journal: Cell Death and Differentiation

Article Title: GLUL mediates FOXO3 O-GlcNAcylation to regulate the osteogenic differentiation of BMSCs and senile osteoporosis

doi: 10.1038/s41418-025-01543-2

Figure Lengend Snippet: A Construction strategies for BMSC-specific Glul- KO ( Glul Prx1 ) mice. B Representative images of Alizarin Red-Alcian Blue double staining of tissues from Glul Prx1 and Glul fl/fl mice( n = 5). C Representative images of Alizarin Red-Alcian Blue double staining of the skull, upper limb, and lower limb from Glul Prx1 and Glul fl/fl mice ( n = 5). D Quantitative analysis of Alizarin Red-Alcian Blue double staining. E Representative images of ARS and ALP staining of BMSCs from Glul Prx1 and Glul fl/fl mice. F Quantitative analysis of ARS and ALP staining. G Representative images of ARS and ALP staining of BMSCs from Glul Prx1 and Glul fl/fl mice after treatment with glutamine. H Quantitative analysis of ARS and ALP staining. I Representative micro-CT images of trabecular bone from the femoral metaphysis of Glul Prx1 and Glul fl/fl mice ( n = 5). J Quantitative analysis of cancellous bone volume (BV/TV, %), trabecular thickness (Tb.Th). K Quantitative analysis of trabecular number (Tb.N), and trabecular separation (Tb.Sp). L Representative images of VON KOSSA staining ( n = 3). M Calcein double-label staining image. N Quantitative analysis of the MAR and BFR/BS in Glul Prx1 and Glul fl/fl mice ( n = 3). ** P < 0.01 vs. other groups; * P < 0.05 vs. other groups.

Article Snippet: Conditional Glul -KO ( Glul fl/fl ) mouse (S-CKO-02655) and Prx1-Cre mouse (C001029) strains were obtained from Cyagen Biotechnology Co., Ltd.

Techniques: Double Staining, Staining, Micro-CT

Journal: Cell Death and Differentiation

Article Title: GLUL mediates FOXO3 O-GlcNAcylation to regulate the osteogenic differentiation of BMSCs and senile osteoporosis

doi: 10.1038/s41418-025-01543-2

Figure Lengend Snippet: A Heatmap of metabolomic assays of the BMSCs from Glul Prx1 and Glul fl/fl mice ( n = 6). B Schematic diagram of the hexosamine biosynthetic pathway. C Heatmap of the proteomics analysis of BMSCs from Glul Prx1 and Glul fl/fl mice ( n = 3). D Western blotting analysis of O-GlcNAcylation levels in BMSCs from Glul Prx1 and Glul fl/fl mice at different osteogenic differentiation timepoints. The numbers on the line graph represent the P -values comparing the relative protein expression levels between the two groups. E Western blotting analysis of UAP1 expression in BMSCs from Glul Prx1 and Glul fl/fl mice at different osteogenic differentiation timepoints. The numbers on the line graph represent the P -values comparing the relative UAP1 expression levels between the two groups. F Western blotting analysis of UAP1 expression in BMSCs from patients in the SOP group ( n = 6) and the control group ( n = 7). G Western blotting analysis of OGT levels in BMSCs from Glul Prx1 and Glul fl/fl mice at different osteogenic differentiation timepoints. H Representative images of ARS and ALP staining of BMSCs from Glul Prx1 and Glul fl/fl mice after transfection with the UAP1 overexpression or control plasmid. ** P < 0.01 vs. other groups; * P < 0.05 vs. other groups.

Article Snippet: Conditional Glul -KO ( Glul fl/fl ) mouse (S-CKO-02655) and Prx1-Cre mouse (C001029) strains were obtained from Cyagen Biotechnology Co., Ltd.

Techniques: Western Blot, Expressing, Control, Staining, Transfection, Over Expression, Plasmid Preparation

Journal: Cell Death and Differentiation

Article Title: GLUL mediates FOXO3 O-GlcNAcylation to regulate the osteogenic differentiation of BMSCs and senile osteoporosis

doi: 10.1038/s41418-025-01543-2

Figure Lengend Snippet: 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.

Article Snippet: Conditional Glul -KO ( Glul fl/fl ) mouse (S-CKO-02655) and Prx1-Cre mouse (C001029) strains were obtained from Cyagen Biotechnology Co., Ltd.

Techniques: Mass Spectrometry, Modification, Residue, Co-Immunoprecipitation Assay, Knock-Out, Mutagenesis, Staining, Transfection, Western Blot, Control, Expressing