Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: THBS1 is up-regulated in the follicular fluid and granulosa cells of PCOS patients and correlates with anti-Müllerian hormone and luteinising hormone. ( A ) Concentrations of THBS1 and IL-6 in follicular fluid from control and PCOS patients were measured by ELISA. individual data points are shown as symbols. ( B ) Relative mRNA expression levels of THBS1 and IL-6 in granulosa cells from control and PCOS patients were determined by qPCR. ( C ) Correlation analyses between follicular fluid THBS1 concentrations and serum AMH and LH levels in PCOS patients. ( D ) Immunofluorescence staining of granulosa cells for FSHR (red) and DAPI (blue). The merged image confirms granulosa cell identity. Scale bar = 50 μm.

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Control, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: AAV9-mediated knockdown of ovarian Thbs1 ameliorates the PCOS-like reproductive phenotype in a DHEA-induced rat model. ( A ) The timeline schematic was created using FigDraw ( https://www.figdraw.com ), illustrating DHEA-induced PCOS establishment and AAV9-mediated THBS1 knockdown intervention. ( B ) Representative HE staining and immunohistochemistry images of ovarian sections. Scale bar = 500 μm. ( C ) Quantification of time spent in each estrous cycle stage. ( D ) Vaginal smear cytology showing estrous cycle stages. Scale bar = 200 μm. ( E ) Serum hormone levels (AMH, FSH, LH, E2, T) measured by ELISA. Different colored bars represent different experimental groups, different symbols indicate individual animal data point. ( F ) Body weight changes during the 28-day protocol. ( G ) Western blot analysis of ovarian THBS1, BAX, and BCL-2 protein expression. ( H ) AAV9-mediated ovarian THBS1 knockdown (37.4% protein reduction, p = 0.006), band intensity was calculated by Image J.

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Knockdown, Staining, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: THBS1 overexpression impairs KGN Cells function. ( A , B ) Validation of THBS1 overexpression by Western blot and qPCR. ( C ) CCK-8 assay showing suppressed cell proliferation over 3 days. ( D , E ) Flow cytometry analysis of apoptosis. ( F , G ) Flow cytometry analysis of cell cycle distribution. ( H – J ) Western blot analysis of apoptosis markers (BAX, BCL-2), inflammatory cytokines (IL-6, TNF-α), steroidogenic enzymes (CYP19, CYP17).

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Over Expression, Biomarker Discovery, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: Molecular docking and 100-ns dynamics simulation reveal a stable association between THBS1 and human latent TGF-1 (PDB 9VJJ). ( A ) Surface representation of THBS1-latent TGF-β1 (PDB 9VJJ) binding interface. Brownish-yellow: TGF-β1, purple-blue: THBS1, red: interfacial binding residues. ( B ) Docking scores and confidence scores of top models. ( C ) Detailed interaction analysis showing hydrogen bonds (blue lines) and salt bridges (yellow dashed lines) between THBS1 (blue) and latent TGF-β1 (PDB 9VJJ) (yellow). ( D ) Per-residue MM/PBSA decomposition. ( E ) RMSD analysis of the protein complex over 100 ns. ( F ) Radius of gyration (Rg) analysis showing structural compactness. ( G ) Solvent-accessible surface area (SASA) analysis. ( H ) Hydrogen bond count throughout the simulation. RMSF analysis of THBS1 ( I ) and latent TGF-β1 (PDB 9VJJ) ( J ) residues. ( K , L ) Gibbs free-energy landscape (FEL) based on RMSD and Rg. ( M ) MM/PBSA binding free energy decomposition.

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Binding Assay, Residue, Solvent

Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: THBS1 overexpression activates the TGF-β1/Smad axis. ( A ) DESeq2 volcano (267 up + 317 down at |log FC| > 1, Padj < 0.05). ( B ) Hierarchical clustering of differentially expressed genes. ( C ) TGF-β/Smad pathway gene expression (28 canonical genes). *, **, *** Data are presented as mean ± SEM. ( D ) Reciprocal co-immunoprecipitation: lanes IgG/IP/Input. ( E ) Active TGF-β1—patient FF. Different colored bars represent different experimental groups, and different symbols indicate individual sample data points. ( F ) Active/Total ratio--patient FF. ( G ) Active TGF-β1—rat serum. ( H ) Active/Total ratio—rat serum. ( I ) Western blot analysis of TGF-β/Smad—KGN cells (THBS1-OE). ( J , K ) Flow cytometry apoptosis (THBS1-OE ± SB-431542). ( L – N ) Western blot analysis of apoptosis markers (BAX, BCL-2), inflammatory cytokines (IL-6, TNF-α), steroidogenic enzymes (CYP19, CYP17), and TGF-β/SMAD2 signaling pathway components.

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Over Expression, Gene Expression, Immunoprecipitation, Western Blot, Flow Cytometry

Journal: Genes & Diseases

Article Title: Mapping intestinal microenvironment heterogeneity in Hirschsprung's disease by spatial transcriptomics: Role of THBS1 in macrophage polarization and neural stem cell regulation

doi: 10.1016/j.gendis.2026.102254

Figure Lengend Snippet: Key gene atlas of extracellular matrix (ECM) regulating the enteric neural crest cells. (A) Expression of ECM-and immune cell-related genes within each cluster. (B) Volcano plots of differentially expressed genes (DEGs) between the spastic and control groups. The DEGs were labeled. p -value < 0.05, Fold change > 1.2. (C) GO enrichment analysis and KEGG pathway analysis of the differentially expressed genes. (D) Violin plots of ECM-related genes in all clusters. The width of a violin plot indicates the kernel density of the expression values. (E) Correlation analyses on DEGs in the cluster with higher ECM scores. Sample sizes: spastic segments ( n = 2), control segments ( n = 1). Statistical significance for DEGs was determined by the Wilcoxon test with p < 0.05 and fold change > 1.2.

Article Snippet: Anti-THBS1 (Proteintech, 18304-1-AP), anti-SDC1 (Abclonal, A4174), or control IgG was added and the lysate was incubated at 4 °C overnight.

Techniques: Expressing, Immune Cell, Control, Labeling, Analysis, Significance Assay