Journal: Bone Research

Article Title: Lumbar instability remodels cartilage endplate to induce intervertebral disc degeneration by recruiting osteoclasts via Hippo-CCL3 signaling

doi: 10.1038/s41413-024-00331-x

Figure Lengend Snippet: Activation of YAP attenuates cartilage degradation during LSI. a Top, representative 3D Micro-CT images of the caudal endplates of L4-5 level (coronal view) from LSI mice with or without Lats-IN-1 treatment. Bottom, quantitative analysis of the total porosity and Tb. Sp. b Representative SOFG staining images of the CEP from LSI mice with or without Lats-IN-1 treatment. c Endplate scores of the caudal endplates based on ( b ). d Left, representative IF staining images of the Collagen II and Collagen X (green) of the CEP from LSI mice with or without Lats-IN-1 treatment. Right, quantitative analysis of the percentage of each area in CEP. e Gene expression of Col2a1 and Col10a1 in CEPCs treated with 12% CTS and Lats-IN-1. f WB analysis of Collgen II, Collgen X and YAP in CEPCs treated with 12% CTS and Lats-IN-1. g , h Left, IF staining of Collagen II and Collagen X in Ctrl, 12% CTS, Ctrl+Lats-IN-1 and 12% CTS+Lats-IN-1 groups. Right, quantitative analysis of the mean fluorescence intensity. F-actin (red), Collagen II and Collagen X (Green), DAPI (blue). * P < 0.05, ** P < 0.01

Article Snippet: YAP agonists Lats-IN-1 (10 μmol/L, MCE) and inhibitor Verteporfin (2 μmol/L, Sigma-Aldrich) was supplemented in culture medium during the entire course of mechanical loading., ,

Techniques: Activation Assay, Micro-CT, Staining, Gene Expression, Fluorescence

Journal: Bone Research

Article Title: Lumbar instability remodels cartilage endplate to induce intervertebral disc degeneration by recruiting osteoclasts via Hippo-CCL3 signaling

doi: 10.1038/s41413-024-00331-x

Figure Lengend Snippet: Loss of YAP triggered CCL3 expression in CEPCs to active osteoclastogenesis. a Volcano plot analysis of the genes expression change profile in CEPCs treated with 12% CTS. b ELISA analysis of CCL3 concentration in the supernatant of CEPCs treated with CTS. c IHC and quantitative analysis of CCL3 in CEP from mice with sham or LSI surgery. d WB analysis of CCL3 in CEPs from mice taken sham or LSI surgery after 8 weeks. e Transwell assay to evaluate the BMM migration under CCL3 attraction. The BMMs were seeded on the upper chamber and different concentrations of CCL3 were added into the lower chamber. Representative images are shown on the left and random cell counting manually in five random fields under ×100 magnification on the right. f WB analysis of YAP and CCL3 in CEPCs treated with 12% CTS and YAP agonists Lats-IN-1. g IF staining of CCL3 in CEPCs treated with 12% CTS and YAP agonists Lats-IN-1. h TRAP staining of BMMs after induction with the supernatant of CTS-treated CEPCs supplemented with CCL3-neutralizing antibody. i Quantative analysis of osteoclast number in ( g ). j Luciferase assay of CCL3 reporter activity to identify the active region of CCL3 promoter in HEK293T cells. k Luciferase assay of CCL3 reporter activity with transfection of control or Yap1-overexpression plasmid in HEK293T cells. l Gene expression analysis of TEADs in CEPCs treated by 12% CTS. * P < 0.05. ** P < 0.01

Article Snippet: YAP agonists Lats-IN-1 (10 μmol/L, MCE) and inhibitor Verteporfin (2 μmol/L, Sigma-Aldrich) was supplemented in culture medium during the entire course of mechanical loading., ,

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transwell Assay, Migration, Cell Counting, Staining, Luciferase, Activity Assay, Transfection, Control, Over Expression, Plasmid Preparation, Gene Expression