Journal: Bioactive Materials

Article Title: From disease model to therapeutic insight: An engineered hydrogel reveals the role of matrix viscous dissipation in intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.021

Figure Lengend Snippet: ITGB1/FAK/YAP mechanotransduction axis mediates the regulation of ECM viscous dissipation on YAP. a, GSEA revealed that “Reactome integrin signaling”, “Gobo focal adhesion assembly”, “Gobo regulation of cytoskeleton” and “Kegg regulation of actin cytoskeleton” were upregulated in the NPCs cultured on V-gel compared with the NPCs cultured on E-gel. b, Heatmap of the series of DEGs related to mechanotransduction. c, Representative IF images of ITGB1 and YAP in NPCs with corresponding quantitative analysis, Scale bars = 50 μm d, Representative immunofluorescence images of p-FAK, Vinculin, YAP, and F-actin with quantitative analysis of cell areas, p-FAK expression and the ratio of nuclear YAP. Relative cell areas and nuclear YAP ratios were quantified across 50 cells from three independent biological replicates (n = 50). Relative fluorescent intensity of p-FAK was quantified from three independent biological replicates (n = 3). e, Western blotting analysis of YAP and p-YAP. All data are presented as means ± SD. Statistical significance was determined by one-way ANOVA with Tukey's multiple-comparisons test. p < 0.05 was considered statistically significant. The symbol “∗” represents a statistical difference.

Article Snippet: During this period, in accordance with a previous study, 10 μL of FAK agonist (ZINC40099027, 10 μM, MCE, HY-134570) or YAP agonist (PY-60, 10 μM, MCE, HY-141644) were injected into the compressed IVDs every other day using a 31G needle.

Techniques: Cell Culture, Immunofluorescence, Expressing, Western Blot

Journal: Bioactive Materials

Article Title: From disease model to therapeutic insight: An engineered hydrogel reveals the role of matrix viscous dissipation in intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.021

Figure Lengend Snippet: Pharmacological activating of YAP alleviates NPCs senescence and IVDD progression. a, Schematic illustration of the in vivo experiments design. Rats received a FAK agonist or a YAP agonist every other day to indirectly or directly activate YAP. b, Images of rats IVDD model c, Representative MRI and X-ray images of the IVDs after injection of FAK agonist and YAP agonist with the corresponding quantitative analysis (n = 5). d, Representative H&E and S. O. staining of IVDs. Scale bars = 1 mm. e, Representative IF images of ACAN and COL II. Scale bars = 100 μm. f, Representative IF images of p16INK4a and YAP. g, Representative IHC images of cGAS with the corresponding quantitative analysis. Scale bars = 100 μm (n = 5). All data are presented as means ± SD. Statistical significance was determined by one-way ANOVA with Tukey's multiple-comparisons test. p < 0.05 was considered statistically significant. The symbol “∗” represents a statistical difference compared to the control group, and the symbol “#” represents a statistical difference compared to the compression group.

Article Snippet: During this period, in accordance with a previous study, 10 μL of FAK agonist (ZINC40099027, 10 μM, MCE, HY-134570) or YAP agonist (PY-60, 10 μM, MCE, HY-141644) were injected into the compressed IVDs every other day using a 31G needle.

Techniques: In Vivo, Injection, Staining, Control

Journal: Bioactive Materials

Article Title: From disease model to therapeutic insight: An engineered hydrogel reveals the role of matrix viscous dissipation in intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.021

Figure Lengend Snippet: Viscous dissipation biomimetic hydrogel alleviates ECM remodeling and NPCs senescence during IVDD. a, Schematic diagram of the mechanical testing procedures. Axial tension-compression and stress-relaxation tests were conducted to evaluate the mechanical properties of IVDs at 12 weeks b, Representative force-displacement curves of all groups. c, Representative stress-relaxation curves of all groups. d, Quantitative analysis of compressive stiffness, tensile stiffness (n = 5). e, Quantitative analysis of normalized NZ and τ 1/2 (n = 5). h, Representative IF images of p16INK4a and YAP at 12 weeks. Scale bars = 50 μm and 20 μm, respectively. i, Representative IHC images of cGAS in NP tissues at 12 weeks. Scale bars = 100 μm (n = 5). The symbol “∗” represents a statistical difference compared to the sham group, and the symbol “#” represents a statistical difference compared to the defect group.

Article Snippet: During this period, in accordance with a previous study, 10 μL of FAK agonist (ZINC40099027, 10 μM, MCE, HY-134570) or YAP agonist (PY-60, 10 μM, MCE, HY-141644) were injected into the compressed IVDs every other day using a 31G needle.

Techniques:

Journal: Frontiers in Oncology

Article Title: Ring finger protein 128 promotes, rather than inhibits, colorectal cancer progression by regulating the Hippo signaling pathway

doi: 10.3389/fonc.2022.1031160

Figure Lengend Snippet: RNF128 ubiquitinates MST for degradation and regulates target genes of the Hippo signaling pathway. (A) Co-IP assays revealed that RNF128 interacts with MST. (B) An IP assay was used to examine the ubiquitination levels of MST. HCT116 cells were treated with MG132 (10 μM) for 6 h after transfection with si-NC and si-RNF128 for 48 h. (C, D) The expression levels of HIF-1α, Bcl-2, p-IκBα and IκBα were examined by western blotting. HCT116 cells were transfected with si-NC and si-RNF128 and treated with verteporfin (1 μM) for 16 h and PY-60 (10 μM) for 24 h. ** P <0.001.

Article Snippet: The proteasome inhibitor MG132 (MedChemExpress, HY-13259) was used to treat HCT116 cells for 6 h, the YAP inhibitor verteporfin (TargetMol, T3112) was used to treat HCT116 cells for 16 h, and the YAP agonist PY-60 (TargetMol, T9566) was used to treat HCT116 cells for 24 h.

Techniques: Co-Immunoprecipitation Assay, Transfection, Expressing, Western Blot

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