integrin αvβ5 inhibitor  (MedChemExpress)

Journal: Frontiers in Immunology

Article Title: Thrombospondin-1 mitigates osteoarthritis progression by inhibiting mechanical stress-induced chondrocyte ferroptosis via the integrin/YAP pathway

doi: 10.3389/fimmu.2025.1577234

Figure Lengend Snippet: The interaction between THBS1 and IntegrinαVβ1 is enhanced under mechanical stress. (A) Interaction network of differential genes, each circle represented a gene, and the size of the circle represented the core level of the gene. The larger the circle, the higher the core level the gene had in the network. (B) Cartoon shows various binding domains of THBS1 and its receptors. (C) Representative immunostaining of THBS1 showing colocalization with integrins αv and β1 (white arrows) but not with integrin β3, CD47, or CD36 under mechanical stress (n = 6). Scale bars, 50 μm. (D) Quantification of colocalization of THBS1 with molecules shown in C using Image J software. (E) Immunoprecipitation (IP) with anti-THBS1 or control immunoglobulin G (IgG) followed by Western blotting for integrins αv and β1 (n = 3). IB, immunoblot. Data were presented as the mean ± SD. **P<0.01.

Article Snippet: IntegrinαVβ1 inhibitor αVβ1 integrin-IN- 1 was obtained from MCE (HY-145363). αVβ1 integrin-IN- 1(100ng/ml) ( ) was added to chondrocytes at the same time of mechanical stress stimulation with or without rhTHBS1, and incubated for 24 hours after the end of mechanical stress stimulation.

Techniques: Binding Assay, Immunostaining, Software, Immunoprecipitation, Control, Western Blot

Journal: Frontiers in Immunology

Article Title: Thrombospondin-1 mitigates osteoarthritis progression by inhibiting mechanical stress-induced chondrocyte ferroptosis via the integrin/YAP pathway

doi: 10.3389/fimmu.2025.1577234

Figure Lengend Snippet: THBS1 inhibits mechanical stress-induced ferroptosis in chondrocytes through integrinαVβ1. Human chondrocytes were stimulated at 1MPa mechanical stress for 2 hours with or without integrinαVβ1 inhibitor αVβ1 integrin-IN- 1(100ng/ml) and rhTHBS1(100ng/ml). Relevant tests were performed 24 hours after the end of mechanical stress stimulation. (A) The cell death ratio of chondrocytes was tested by cell death/live analysis. Scale bar = 50 μm. (B) The cell number of PI (red fluorescence)/calcein (green fluorescence) reflected the cell death ratio (n=3 for each group). (C) Mitochondrial membrane potential was detected by JC-1 assay. Scale bar = 10 μm. (D) The relative IOD ratio of red fluorescence to green fluorescence was used for quantitative analysis (n=3 for each group). (E) Representative TEM images of chondrocytes of indicated groups. (n=3 for each group). Green arrows show the normal mitochondria. Red arrows show the shrunken mitochondria. Scale bars, 1 μm (low field), 500 nm (high field). (F) Representative fluorescence images of mitochondria in chondrocytes. Scale bar = 50 μm. (G) Quantitative analysis of fluorescence intensity (n=3 for each group). (H) Representative images of ROS levels in chondrocytes. Scale bar = 50 μm. (I) Quantitative analysis of fluorescence intensity (n=3 for each group). (J) The expression of GSH in chondrocytes of indicated groups was detected by ELISA (n=3 for each group). (K) Western blot (WB) analysis of Col2, MMP-9 and GPX4. (L-N) Quantification of WB analysis (n=3 for each group). Data were presented as the mean ± SD. **P<0.01.

Article Snippet: IntegrinαVβ1 inhibitor αVβ1 integrin-IN- 1 was obtained from MCE (HY-145363). αVβ1 integrin-IN- 1(100ng/ml) ( ) was added to chondrocytes at the same time of mechanical stress stimulation with or without rhTHBS1, and incubated for 24 hours after the end of mechanical stress stimulation.

Techniques: Fluorescence, Membrane, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Immunology

Article Title: Thrombospondin-1 mitigates osteoarthritis progression by inhibiting mechanical stress-induced chondrocyte ferroptosis via the integrin/YAP pathway

doi: 10.3389/fimmu.2025.1577234

Figure Lengend Snippet: THBS1 inhibits chondrocyte ferroptosis through the integrinαVβ1/YAP pathway. THBS1 knockout chondrocytes and normal chondrocytes were subjected to 1 MPa mechanical stress for 2 hours, with or without the addition of YAP1 inhibitor PROTAC YAP d1 (20μM) and rhTHBS1(100ng/ml). Immediately following stimulation, IF detection was performed. Additional analyses were conducted after chondrocytes were further incubated with or without YAP1 inhibitor PROTAC YAP d1 (20μM) and rhTHBS1(100ng/ml) for 24 hours post-stimulation. (A) Representative image of chondrocytes undergoing IF detection immediately after mechanical stress stimulation. YAP (red) and DAPI (blue) are also shown. Scale bar = 20 μm. (B) Quantification of YAP localization. In each experiment 150 to 200 cells were evaluated (n = 3). YAP localization in the nucleus (green) or cytoplasm (red) is shown. (C) Western blot (WB) analysis of YAP1. (D) Quantification of WB analysis (n=3 for each group). (E) Representative image of chondrocytes undergoing IF detection immediately after mechanical stress stimulation. YAP (red) and DAPI (blue) are also shown. Scale bar = 20 μm. (F) Quantification of YAP localization. In each experiment 150 to 200 cells were evaluated (n = 3). (G) Western blot (WB) analysis of YAP1. (H) Quantification of WB analysis (n=3 for each group). (I) Mitochondrial membrane potential was detected by JC-1 assay. Scale bar = 10 μm. (J) The relative IOD ratio of red fluorescence to green fluorescence was used for quantitative analysis (n=3 for each group). (K) Representative TEM images of chondrocytes of indicated groups. (n=3 for each group). Green arrows show the normal mitochondria. Red arrows show the shrunken mitochondria. Scale bars, 5.0 μm (low field), 500 nm (high field). (L) Representative fluorescence images of mitochondria in chondrocytes. Scale bar = 50 μm. (M) Quantitative analysis of fluorescence intensity (n=3 for each group). (N) The expression of GSH in chondrocytes of indicated groups was detected by ELISA (n=3 for each group). Data were presented as the mean ± SD. NS P>0.05 *P<0.05, **P<0.01.

Article Snippet: IntegrinαVβ1 inhibitor αVβ1 integrin-IN- 1 was obtained from MCE (HY-145363). αVβ1 integrin-IN- 1(100ng/ml) ( ) was added to chondrocytes at the same time of mechanical stress stimulation with or without rhTHBS1, and incubated for 24 hours after the end of mechanical stress stimulation.

Techniques: Knock-Out, Incubation, Western Blot, Membrane, Fluorescence, Expressing, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: CAF-derived POSTN induces the translation to EMT-subtype via PI3K/AKT/β-catenin signaling in PDAC Cells. (A) Differential gene expression analysis of ductal cells based on fibroblast-derived POSTN levels. Tumor samples from the CRA001160 dataset were classified into high- and low- POSTN groups (75th percentile cutoff), and ductal cell gene expression profiles were compared to identify differences linked to CAF-derived POSTN. (B) GSEA of upregulated genes in ductal cells from POSTN-high samples identified enriched KEGG pathways. (C-F) GSEA of upregulated genes revealed enrichment in the EMT pathway, focal adhesion pathway, PI3K-AKT pathway, and WNT pathway. (G-H) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment with rhPOSTN at varying concentrations. (I) Schematic illustration of how CAF-derived POSTN drives the EMT phenotype in PDAC cells via integrin αvβ5/FAK/PI3K/AKT/β-catenin signaling pathway.

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Derivative Assay, Gene Expression, Western Blot, Expressing, Phospho-proteomics

Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: Integrin αvβ5 inhibitors partially reverse POSTN-induced proliferation, colony formation, migration, and invasion of PDAC cells. (A) Co-localization of POSTN and intergrin β5 in PDAC tissues. Immunofluorescence staining showing POSTN (green), integrin β5 (red), and nuclei (blue) in PDAC patient resection specimens. Scale bars, 25 μm. (B-C) Effect of integrin αvβ5 inhibition on POSTN-induced proliferation in BxPC-3 and PANC-1 cells. Cells were treated with: (1) negative control, (2) 500 ng/mL rhPOSTN, (3) integrin αvβ5 inhibitor (HY-16141) at 1/5 IC 50 concentration, or (4) integrin αvβ5 inhibitor pretreated for 24 hours, followed by 500 ng/mL rhPOSTN. Cell proliferation was assessed using CCK-8 assays. (D) Colony formation assays in BxPC-3 and PANC-1 cells following the same treatments as in (B-C). (E-F) Wound healing assays in BxPC-3 and PANC-1 cells after treatments as described in (B-C), assessing migration capacity. (G-H) Transwell assays in BxPC-3 and PANC-1 cells to assess migation (G) and invasion (H) under the same treatment as in (B-C). (I) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment as described in (B-C).

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Migration, Immunofluorescence, Staining, Inhibition, Negative Control, Concentration Assay, CCK-8 Assay, Western Blot, Expressing, Phospho-proteomics