Journal: Molecular Medicine

Article Title: Targeting casein kinase 2 and ubiquitin-specific protease 7 to modulate RUNX2-mediated osteogenesis in chronic kidney disease

doi: 10.1186/s10020-025-01222-5

Figure Lengend Snippet: In vitro Cell Experimental Validation of CK2 and RUNX2/USP7 Interaction. Note: ( A ) Western Blot analysis of the protein expression levels of CK2 and RUNX2 in various groups of MC3T3-E1 cells; ( B ) Western Blot analysis of the protein expression levels of RUNX2 in different groups of MC3T3-E1 cells; ( C ) RT-qPCR analysis of the relative expression levels of osteoblast marker genes (ALP, Collagen-1, and Osteocalcin) in different groups of MC3T3-E1 cells; ( D ) Immunoprecipitation experiment to detect the interaction between RUNX2 and CK2; ( E ) Phosphorylation kinase assay assessing the regulatory role of CK2 in the phosphorylation of RUNX2; ( F ) Ubiquitination experiment measuring the ubiquitination levels of RUNX2 in various groups of cells after treatment with MG132; ( G ) In vitro cell experimental validation of the mutual regulation among CK2, USP7, and RUNX2. * indicates a significant difference between the groups ( P < 0.05), and all cellular experiments were conducted in triplicate

Article Snippet: The membrane was blocked with 5% BSA at room temperature for 1 h and then incubated overnight at 4 °C with primary antibodies against rabbit polyclonal antibodies for RUNX2 (ab236639; Abcam, Cambridge, UK), CK2 (10992-1-AP; Proteintech, USA), USP7 (ab108931; Abcam, Cambridge, UK), and GAPDH (ab108931; Abcam, Cambridge, UK), with β-Actin used as an internal control.

Techniques: In Vitro, Biomarker Discovery, Western Blot, Expressing, Quantitative RT-PCR, Marker, Immunoprecipitation, Phospho-proteomics, Kinase Assay, Ubiquitin Proteomics

Journal: Molecular Medicine

Article Title: Targeting casein kinase 2 and ubiquitin-specific protease 7 to modulate RUNX2-mediated osteogenesis in chronic kidney disease

doi: 10.1186/s10020-025-01222-5

Figure Lengend Snippet: Impact of CK2 Silencing on RUNX2 Expression and Bone Turnover Rate, Bone Density, and Bone Strength in Mice. Note: ( A ) RT-qPCR to detect RUNX2 expression levels in various tissues; ( B ) Western blot analysis of CK2 and RUNX2 protein levels in bone tissue of each group of mice; ( C ) ELISA assessment of PINP and CTx expression levels in the serum of each group of mice; ( D ) Micro-CT 3D scanning (200 μm) of trabecular and cortical bone regions in each group of mice; ( E ) Quantitative evaluation of bone parameters from the Micro-CT 3D scanning results, including BV/TV, Tb. N (mm − 1 ), Tb. Th (mm), Tb. Sp, mm, Ct. Ar (mm 2 ), and Ct. Th (mm 2 ); ( F ) H&E staining of tibial bone tissue in each group of mice (200 μm); ( G ) Quantitative histomorphological assessment of the MASSON stained tissues. *indicates significance in comparisons between the two groups ( P < 0.05), and ** denotes significance at P < 0.01. Each group consisted of 6 mice

Article Snippet: The membrane was blocked with 5% BSA at room temperature for 1 h and then incubated overnight at 4 °C with primary antibodies against rabbit polyclonal antibodies for RUNX2 (ab236639; Abcam, Cambridge, UK), CK2 (10992-1-AP; Proteintech, USA), USP7 (ab108931; Abcam, Cambridge, UK), and GAPDH (ab108931; Abcam, Cambridge, UK), with β-Actin used as an internal control.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Micro-CT, Staining

Journal: Molecular Medicine

Article Title: Targeting casein kinase 2 and ubiquitin-specific protease 7 to modulate RUNX2-mediated osteogenesis in chronic kidney disease

doi: 10.1186/s10020-025-01222-5

Figure Lengend Snippet: The Impact of CK2 Regulating RUNX2 on Bone Turnover Rate, Bone Density, and Bone Strength in CKD-MBD Mice. Note: ( A ) RT-qPCR to detect RUNX2 expression levels in various tissues; ( B ) Western blot analysis of CK2 and RUNX2 protein levels in bone tissue of each group of mice; ( C ) ELISA assessment of PINP and CTx expression levels in the serum of each group of mice; ( D ) Micro-CT 3D scanning (200 μm) of trabecular and cortical bone regions in each group of mice; ( E ) Quantitative evaluation of bone parameters from the Micro-CT 3D scanning results, including BV/TV, Tb. N (mm − 1 ), Tb. Th (mm), Tb. Sp, mm, Ct. Ar (mm 2 ), and Ct. Th (mm 2 ); ( F ) H&E staining of tibial bone tissue in each group of mice (200 μm); ( G ) Quantitative histomorphological assessment of the MASSON stained tissues. * indicates significance in comparisons between the two groups ( P < 0.05). Each group comprised 6 mice.=

Article Snippet: The membrane was blocked with 5% BSA at room temperature for 1 h and then incubated overnight at 4 °C with primary antibodies against rabbit polyclonal antibodies for RUNX2 (ab236639; Abcam, Cambridge, UK), CK2 (10992-1-AP; Proteintech, USA), USP7 (ab108931; Abcam, Cambridge, UK), and GAPDH (ab108931; Abcam, Cambridge, UK), with β-Actin used as an internal control.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Micro-CT, Staining

Journal: Molecular Medicine

Article Title: Targeting casein kinase 2 and ubiquitin-specific protease 7 to modulate RUNX2-mediated osteogenesis in chronic kidney disease

doi: 10.1186/s10020-025-01222-5

Figure Lengend Snippet: Molecular Mechanism Diagram of CK2 Phosphorylation Modifications on RUNX2 Affecting MBD in CKD Mice

Article Snippet: The membrane was blocked with 5% BSA at room temperature for 1 h and then incubated overnight at 4 °C with primary antibodies against rabbit polyclonal antibodies for RUNX2 (ab236639; Abcam, Cambridge, UK), CK2 (10992-1-AP; Proteintech, USA), USP7 (ab108931; Abcam, Cambridge, UK), and GAPDH (ab108931; Abcam, Cambridge, UK), with β-Actin used as an internal control.

Techniques: Phospho-proteomics