Journal: Cell Death Discovery

Article Title: Glycolytic reprogramming mediated by the ADAM12/IGF1 axis promotes ossification of the posterior longitudinal ligament

doi: 10.1038/s41420-026-03044-8

Figure Lengend Snippet: A Representative immunofluorescence images and quantification of glycolytic enzymes PKM2 and LDHA in human PLL and OPLL tissues. Scale bar = 100 µm, n = 3. B Western blot analysis of GLUT1, PKM2, and LDHA protein levels in PLL and OPLL cells. n = 3. C , D . Glucose uptake and lactate production in PLL and OPLL cells. n = 3. E ECAR measured by Seahorse XF Analyzer to assess the glycolytic capacity of PLL and OPLL cells. n = 3. F Representative images showing TMRE staining (orange-red) in PLL and OPLL cells. The bar graph depicts the quantification of relative TMRE fluorescence intensity. Scale bar = 30 µm, n = 3. G Western blot analysis of PDK1, Cyto c, and ATP5A in OPLL and PLL cells. H Intracellular ROS levels measured by DCFH-DA fluorescence. Scale bar = 50 µm. I OCR measured by Seahorse XF Analyzer to assess mitochondrial respiration. n = 3. J Western blot analysis of RUNX2, OSX, and ALP during a 15day time course of osteogenic differentiation of ligament cells. K Western blot analysis of GLUT1, HK2, and LDHA during a 15day time course of osteogenic differentiation of ligament cells. L Western blot analysis of PDK1, Cyto c, and ATP5A during a 15day time course of osteogenic differentiation of ligament cells. Data are presented as mean ± standard deviation.

Article Snippet: The primary antibodies and dilutions used were: ADAM12 (Proteintech, 14139-1-AP, 1:100), RUNX2 (Abclonal, A11753, 1:600), OCN (Proteintech, 23418-1-IG, 1:600), PKM2 (Proteintech, 60268-1-IG, 1:1500), LDHA (Servicebio, GB11342-100, 1:300), CD31 (Servicebio, GB11063-2-50, 1:1500), PRG4 (Abcam, ab28484, 1:200) and Vimentin (Proteintech, 10366-1-AP, 1:200).

Techniques: Immunofluorescence, Western Blot, Staining, Fluorescence, Standard Deviation

Journal: Cell Death Discovery

Article Title: Glycolytic reprogramming mediated by the ADAM12/IGF1 axis promotes ossification of the posterior longitudinal ligament

doi: 10.1038/s41420-026-03044-8

Figure Lengend Snippet: A Venn diagram illustrating the overlap of hub genes identified from the PPI network using five different centrality algorithms: MCC, DMNC, EPC, MNC, and Degree. B Key genes identified by the DMNC method within the PPI network. C Correlation analysis between the glycolysis flux score and ADAM12 expression in OPLL and PLL tissues. D Distribution of ADAM12 across the major cell types in posterior longitudinal ligament tissue. E Scatter plot showing the positive correlation between ADAM12 expression and the glucose metabolism pathway activity score across individual ligament cells. F Representative immunofluorescence images and quantification of ADAM12 expression in human OPLL and PLL tissues. Scale bar = 50 μm, n = 3. G Representative immunofluorescence images showing co-localization of ADAM12 (green) and PKM2 (red) in OPLL tissue. Scale bar = 30 μm. H , I Quantitative PCR analysis of ADAM12L and ADAM12S mRNA levels in PLL and OPLL cells. n = 3. J Quantitative PCR analysis of the expression levels of the ADAM12L and ADAM12S isoforms in OPLL cells. n = 3. K Western blot analysis of ADAM12 protein levels in PLL and OPLL cells. n = 3. L ELISA analysis of ADAM12 protein levels in PLL and OPLL ligament cells supernatant. n = 5. Data are presented as mean ± standard deviation.

Article Snippet: The primary antibodies and dilutions used were: ADAM12 (Proteintech, 14139-1-AP, 1:100), RUNX2 (Abclonal, A11753, 1:600), OCN (Proteintech, 23418-1-IG, 1:600), PKM2 (Proteintech, 60268-1-IG, 1:1500), LDHA (Servicebio, GB11342-100, 1:300), CD31 (Servicebio, GB11063-2-50, 1:1500), PRG4 (Abcam, ab28484, 1:200) and Vimentin (Proteintech, 10366-1-AP, 1:200).

Techniques: Expressing, Activity Assay, Immunofluorescence, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Cell Death Discovery

Article Title: Glycolytic reprogramming mediated by the ADAM12/IGF1 axis promotes ossification of the posterior longitudinal ligament

doi: 10.1038/s41420-026-03044-8

Figure Lengend Snippet: A Heatmap showing the relative abundance of central carbon metabolites in control (NC), osteogenically differentiated (NC-OD), and ADAM12-overexpressing osteogenically differentiated (OE-OD) ligament cells. n = 3. B Box plots quantifying the levels of key glycolytic intermediates (fructose-1,6-bisphosphate, 3-phosphoglycerate, pyruvate, and lactate) from the metabolomics data. n = 3. C , D Western blot analysis of glycolytic markers (GLUT1, HK2, PKM2, LDHA) and mitochondrial markers (PGC1α, mtTFA, ATP5A, cytochrome c) in ligament cells after ADAM12 knockdown and overexpression. n = 3. E , F Glucose uptake capacity in ligament cells after ADAM12 knockdown and overexpression. n = 3. G , H Lactate production in ligament cells after ADAM12 knockdown and overexpression. n = 3. I , J Intracellular pyruvate content in ligament cells after ADAM12 knockdown and overexpression. n = 3. K , L ECAR measured by Seahorse XF Analyzer in ligament cells after ADAM12 knockdown and overexpression. n = 3. Data are presented as mean ± standard deviation.

Article Snippet: The primary antibodies and dilutions used were: ADAM12 (Proteintech, 14139-1-AP, 1:100), RUNX2 (Abclonal, A11753, 1:600), OCN (Proteintech, 23418-1-IG, 1:600), PKM2 (Proteintech, 60268-1-IG, 1:1500), LDHA (Servicebio, GB11342-100, 1:300), CD31 (Servicebio, GB11063-2-50, 1:1500), PRG4 (Abcam, ab28484, 1:200) and Vimentin (Proteintech, 10366-1-AP, 1:200).

Techniques: Control, Western Blot, Knockdown, Over Expression, Standard Deviation