adamts1  (Bioss)

Journal: Animals : an Open Access Journal from MDPI

Article Title: Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

doi: 10.3390/ani15162354

Figure Lengend Snippet: Predominant localization of ADAMTS1 in follicular granulosa cells of the ovine ovary. ( A ) Hematoxylin and eosin (HE) staining alongside immunohistochemical detection of ADAMTS1 in ovarian tissue. The left panel illustrates HE staining, while the right panel displays ADAMTS1 immunostaining (bar = 50 μm). ( B ) Immunofluorescence analysis of granulosa cells and oocytes. GC-NC: granulosa cells with negative control staining; GC-FSHR: granulosa cells immunostained for FSHR; GC-ADAMTS1: granulosa cells immunostained for ADAMTS1; OC-ADAMTS1: oocytes immunostained for ADAMTS1 (bar = 100 μm).

Article Snippet: Following 24 h culture, cells were fixed in 4% tissue fixative for 30 min, then permeabilized with 0.2% Triton X-100 (Coolaber, Beijing, China) for 20 min. Non-specific binding sites were blocked by treatment with 5% bovine serum albumin (BSA) (Servicebio, Wuhan, China) for 3 h. The cells were subsequently incubated overnight at 4 °C with primary antibodies targeting FSHR and ADAMTS1 (Bioss, Beijing, China).

Techniques: Staining, Immunohistochemical staining, Immunostaining, Immunofluorescence, Negative Control

Journal: Animals : an Open Access Journal from MDPI

Article Title: Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

doi: 10.3390/ani15162354

Figure Lengend Snippet: Design and validation of ADAMTS1 knockdown and overexpression systems in ovarian granulosa cells. ( A ) Four siRNA targeting sequences against ADAMTS1 were designed using an online prediction tool. ( B ) Knockdown efficiency was verified by quantifying ADAMTS1 mRNA and protein levels post-transfection in granulosa cells. Among the four siRNA constructs tested, si-ADAMTS1-2 demonstrated the most robust knockdown efficiency, significantly reducing both ADAMTS1 mRNA and protein expression levels in ovarian granulosa cells. ( C ) Agarose gel electrophoresis confirming successful PCR amplification of the ADAMTS1 coding sequence (CDS). Using the DL10,000 DNA marker, agarose gel electrophoresis confirmed successful amplification of the ADAMTS1 coding sequence (CDS), with the observed band (2903 bp) migrating between 2000 and 4000 bp, consistent with the expected product size. ( D ) Restriction enzyme digestion analysis using EcoR I and BamH I to verify linearized pcDNA3.1-EGFP vector and EGFP-ADAMTS1 recombinant plasmid. Using the DL10000 DNA marker, the linearized pcDNA3.1-EGFP vector migrated as a single band between 7000 and 10,000 bp, consistent with its expected size of 6750 bp. For the EGFP-ADAMTS1 recombinant plasmid, digestion with EcoRI and BamHI yielded fragments of 6750 bp, 994 bp, 900 bp, 475 bp, 387 bp, and 151 bp, all of which matched their predicted sizes by electrophoretic mobility. ( E ) Schematic representation of the ADAMTS1 overexpression vector construct. The size of the overexpression vector EGFP-ADAMTS1 is 9657 bp. ( F ) Overexpression efficacy was assessed by measuring ADAMTS1 transcriptional and translational levels following plasmid transfection. ( G ) Sanger sequencing chromatogram confirming the accurate insertion of EGFP-ADAMTS1 in the expression vector. * for p < 0.05; ** for p < 0.01.

Article Snippet: Following 24 h culture, cells were fixed in 4% tissue fixative for 30 min, then permeabilized with 0.2% Triton X-100 (Coolaber, Beijing, China) for 20 min. Non-specific binding sites were blocked by treatment with 5% bovine serum albumin (BSA) (Servicebio, Wuhan, China) for 3 h. The cells were subsequently incubated overnight at 4 °C with primary antibodies targeting FSHR and ADAMTS1 (Bioss, Beijing, China).

Techniques: Biomarker Discovery, Knockdown, Over Expression, Transfection, Construct, Expressing, Agarose Gel Electrophoresis, Amplification, Sequencing, Marker, Plasmid Preparation, Recombinant

Journal: Animals : an Open Access Journal from MDPI

Article Title: Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

doi: 10.3390/ani15162354

Figure Lengend Snippet: ADAMTS1 enhances proliferation in follicular granulosa cells. ( A ) Morphological observations of granulosa cells at 0, 24, and 48 h post ADAMTS1 knockdown and overexpression. ADAMTS1 knockdown significantly reduced cell numbers, whereas ADAMTS1 overexpression markedly increased cell numbers and enhanced EGFP fluorescence intensity in transfected ovarian granulosa cells. ( B ) Quantification of granulosa cell numbers at 0, 24, and 48 h following modulation of ADAMTS1 expression. At 48 h post-transfection, ADAMTS1 knockdown significantly reduced granulosa cell numbers by 40.0% ( p < 0.01), while its overexpression enhanced cellular proliferation by 1.13-fold ( p < 0.01). ( C ) Flow cytometric analysis of apoptosis in granulosa cells subjected to ADAMTS1 silencing and overexpression. Relative to control groups, ADAMTS1 knockdown reduced the proliferation rate of EdU-positive granulosa cells (GCs) to 76.3% of baseline levels ( p < 0.01), whereas ADAMTS1 overexpression enhanced proliferative activity to 148.2% ( p < 0.01). ( D ) After knockdown of ADAMTS1 , the mRNA and protein expression levels of Bcl2 ( p < 0.05), Bax ( p < 0.05), and caspase3 ( p < 0.05) in the granulosa cells were significantly increased. ( E ) After overexpression of ADAMTS1 , the mRNA and protein expression levels of Bcl2 ( p < 0.05), Bax ( p < 0.05), and caspase3 ( p < 0.05) in the granulosa cells were significantly decreased. ns for not significant; * for p < 0.05; ** for p < 0.01.

Article Snippet: Following 24 h culture, cells were fixed in 4% tissue fixative for 30 min, then permeabilized with 0.2% Triton X-100 (Coolaber, Beijing, China) for 20 min. Non-specific binding sites were blocked by treatment with 5% bovine serum albumin (BSA) (Servicebio, Wuhan, China) for 3 h. The cells were subsequently incubated overnight at 4 °C with primary antibodies targeting FSHR and ADAMTS1 (Bioss, Beijing, China).

Techniques: Knockdown, Over Expression, Fluorescence, Transfection, Expressing, Control, Activity Assay

Journal: Animals : an Open Access Journal from MDPI

Article Title: Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

doi: 10.3390/ani15162354

Figure Lengend Snippet: Summary of RNA sequencing results and functional enrichment analysis of differentially expressed genes (DEGs). ( A ) Quantification of upregulated and downregulated DEGs in ADAMTS1 knockdown and overexpression groups. ( B ) Volcano plot illustrating DEGs identified between the ADAMTS1 knockdown group and control. Transcriptomic analysis identified 2684 differentially expressed genes (DEGs), comprising 159 upregulated (5.9%) and 2,525 downregulated (94.1%) transcripts, based on the established cutoff criteria ( p < 0.05, |log2 fold change| ≥ 1). ( C ) Volcano plot depicting DEGs between the ADAMTS1 overexpression group and control. Transcriptomic analysis identified 459 differentially expressed genes (DEGs), comprising 271 upregulated (59.0%) and 188 downregulated (41.0%) transcripts, based on the established cutoff criteria ( p < 0.05, |log2 fold change| ≥ 1). ( D ) Gene Ontology (GO) enrichment analysis for DEGs comparing the ADAMTS1 knockdown group with control. ( E ) GO enrichment analysis for DEGs between the ADAMTS1 overexpression group and control. ( F ) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for DEGs in the ADAMTS1 knockdown versus control comparison. ( G ) KEGG pathway enrichment for DEGs in the ADAMTS1 overexpression versus control comparison.

Article Snippet: Following 24 h culture, cells were fixed in 4% tissue fixative for 30 min, then permeabilized with 0.2% Triton X-100 (Coolaber, Beijing, China) for 20 min. Non-specific binding sites were blocked by treatment with 5% bovine serum albumin (BSA) (Servicebio, Wuhan, China) for 3 h. The cells were subsequently incubated overnight at 4 °C with primary antibodies targeting FSHR and ADAMTS1 (Bioss, Beijing, China).

Techniques: RNA Sequencing, Functional Assay, Knockdown, Over Expression, Control, Comparison

Journal: Animals : an Open Access Journal from MDPI

Article Title: Transcriptome Analysis Reveals the Molecular Mechanisms by Which ADAMTS1 Influences the Proliferation of Ovarian Granulosa Cells in Sheep

doi: 10.3390/ani15162354

Figure Lengend Snippet: Functional characterization of DEGs. ( A ) Venn diagram illustrating the overlap of DEGs across four groups: downregulated and upregulated genes following ADAMTS1 knockdown, and downregulated and upregulated genes following ADAMTS1 overexpression. Through comparative analysis, it was found that in all comparison groups, there were 17 differentially expressed genes ( p < 0.05, |log2 fold change| ≥ 1). ( B ) Detailed gene information for the 17 co-expressed DEGs (co-DEGs). ( C ) Heatmap depicting the expression patterns of the 17 co-DEGs in the ADAMTS1 knockdown group. ( D ) Heatmap illustrating the expression profiles of the 17 co-DEGs in the ADAMTS1 overexpression group. ( E ) Protein-protein molecular docking analysis between ADAMTS1 (colored blue) and PSAT1 (colored green). ( F ) Predicted interaction interface highlighting critical amino acid residues involved in binding: ADAMTS1 key residues include Ser419, Ser440, Gln441, Asp438, Lys294, Ile541, and Asp530; PSAT1 key residues include Arg336, Asn9, Ala15, His19, Gln26, and Ser38. ( G ) The mRNA expression levels of PSAT1 and SLC6A9 in the ADAMTS1 knockdown group and the overexpression group. ( H ) The protein expression levels of PSAT1 in the ADAMTS1 knockdown group and the overexpression group. * for p < 0.05; ** for p < 0.01.

Article Snippet: Following 24 h culture, cells were fixed in 4% tissue fixative for 30 min, then permeabilized with 0.2% Triton X-100 (Coolaber, Beijing, China) for 20 min. Non-specific binding sites were blocked by treatment with 5% bovine serum albumin (BSA) (Servicebio, Wuhan, China) for 3 h. The cells were subsequently incubated overnight at 4 °C with primary antibodies targeting FSHR and ADAMTS1 (Bioss, Beijing, China).

Techniques: Functional Assay, Knockdown, Over Expression, Comparison, Expressing, Binding Assay

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 levels were increased in the serum of patients with myocardial fibrosis, in the hearts of mice after CFPMI and in cardiac fibroblasts stimulated with fibrotic factors. A . ADAMTS1 levels in clinical serum samples were evaluated via ELISA. N = 30. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. B . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. C . Survival rate of mice in each group. D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E and F . ADAMTS1 expression in mouse heart tissue was determined through Western blot and IHC. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 12, 24, or 48 h to construct in vitro models. The cells were divided into four groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups. G and H . Western blot detection of the expression of ADAMTS1, Collagen I and FN. I. The expression levels of ADAMTS1, Collagen I and FN were measured via qRT‒PCR. After the optimal time point (48 h) was determined, the mice were further divided into the control, TGF-β1 and Ang II groups. J . ADAMTS1 expression in cells was assessed by IF. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 15, 30, 60, 90, and 120 min. K. qRT-PCR detection of the mRNA expression of ADAMTS1, Collagen I and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Construct, Staining, Expressing, Western Blot, In Vitro, Control, Quantitative RT-PCR

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: SMAD2 regulated ADAMTS1 expression in human and mouse cardiac fibroblasts induced by TGF-β1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 15, 30, 60, 90, and 120 min. A and B . Western blot analysis of the expression of SMAD2, p-SMAD2 and ADAMTS1. Human and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 for 48 h, and the cells were divided into Control and TGF-β1 groups. C . Human and mouse cardiac fibroblasts were induced with 10 ng/mL TGF-β1 for 12 h, 24 h, 48 h, and 72 h. qRT‒PCR was performed to detect the mRNA levels of ADAMTS1, Collagen I, and FN. D . IF staining was conducted to determine the expression of p-SMAD2 and ADAMTS1 at 48 h. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. E . p-SMAD2 expression in mouse heart tissue was measured by IHC. F . ChIP verification of the interaction between SMAD2 and ADAMTS1. Then, we interfered with SMAD2 expression. The groups were the si-NC, si-SMAD2-1, and si-SMAD2-2 groups. G . SMAD2 expression was assessed via qRT‒PCR. After the best si-SMAD2 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-SMAD2, and TGF-β1 + si-SMAD2 groups. H . Western blot detection of ADAMTS1, Collagen I, and FN expression in cells. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Expressing, Western Blot, Control, Staining, Construct

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Overexpression of ADAMTS1 enhanced the production of collagen fiber proteins in human and mouse cardiac fibroblasts induced by TGF-β1. We overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1 (48 h), oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. A . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. B-D . IF staining of Collagen I and FN expression. Additionally, we interfered with ADAMTS1; the cells were grouped as follows: si-NC, si-ADAMTS1-1, and si-ADAMTS1-2. E . ADAMTS1 expression was determined through qRT‒PCR. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. F and G . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. H and I . IF staining of Collagen I and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Over Expression, Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Knockdown of ADAMTS1 alleviated TGF-β1-induced fibrosis by downregulating HDAC6 protein expression. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. A . HDAC6 expression in mouse heart tissue was determined through IHC. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC and si-ADAMTS1 groups. B . ADAMTS1 and HDAC6 expression was assessed via qRT‒PCR. C . Western blot analysis of ADAMTS1 and HDAC6 expression. Furthermore, we overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1, oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. D . Western blot detection of HDAC6 expression. After the si-ADAMTS1 group was selected, the cells were further divided into the si-NC, TGF-β1, si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. E. Western blot detection of HDAC6 expression. Moreover, cells were treated with the HDAC6 inhibitor ACY1215 (5 μM) for 48 h and then divided into the si-NC, si-ADAMTS1, si-NC + ACY1215, and si-ADAMTS1 + ACY1215 groups. F. Western blot detection of TGF-β1, Collagen I, FN, and ADAMTS1 expression. Subsequently, experiments were performed in human and mouse cardiac fibroblasts with ADAMTS1 overexpression followed by treatment with the HDAC6 inhibitor ACY1215. G. Western blot analysis was conducted to detect the expression levels of ADAMTS1 and HDAC6. H. Western blot analysis was performed to determine the expression levels of TGF-β1, Collagen I, and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Knockdown, Expressing, Construct, Western Blot, Over Expression

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 interacted with HDAC6 during fibrosis. A . Co-IP verification of the interaction of ADAMTS1 with HDAC6, with ADAMTS1 as the bait protein. In the experiment, we first captured the ADAMTS1 protein using a specific antibody and then used Co-IP technology to detect its interaction with HDAC6. B . Co-IP verification of the interaction of HDAC6 with ADAMTS1, with HDAC6 as the bait protein. In the experiment, we captured the HDAC6 protein using a specific antibody and then used Co-IP technology to detect its interaction with ADAMTS1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 48 h, and the cells were further divided into control and TGF-β1 groups. C . IF staining was performed to evaluate the colocalization of ADAMTS1 and HDAC6 in TGF-β1-treated human and mouse cardiac fibroblasts. D . Changes in the level of ubiquitinated HDAC6 protein in human and mouse cardiac fibroblasts transfected with oe-ADAMTS1/si-ADAMTS1 or oe-NC/si-NC in the presence of 10 μM MG132. E. In si-ADAMTS1-treated human and mouse cardiac fibroblasts, MG132 (10 μM) was added to detect the ubiquitination levels of ADAMTS1 protein. F. Human and mouse cardiac fibroblasts were treated with TGF-β1 and subjected to SMAD2 knockdown to determine the ubiquitination levels of ADAMTS1 protein. N = 3. *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Co-Immunoprecipitation Assay, Control, Staining, Transfection, Ubiquitin Proteomics, Knockdown

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-ADAMTS1 treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, and CFPMI + sh-ADAMTS1 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. D . HDAC6 expression was determined through qRT‒PCR. E. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. F. Western blot detection of Collagen I, and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: shRNA, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-HDAC6 transfection combined with ADAMTS1 inhibitor treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, CFPMI + anti-ADAMTS1, CFPMI + sh-HDAC6, and CFPMI + anti-ADAMTS1 + sh-HDAC6 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C and D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E. HDAC6 expression was determined through qRT‒PCR. F. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. G. Western blot detection of Collagen I and FN expression. N = 3. ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: shRNA, Transfection, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: TGF-β1/SMAD2 regulated ADAMTS1 by mediating CFPMI through HDAC6 ubiquitination

Article Snippet: Primary antibodies against ADAMTS1 (12,749–1-AP, 1:200, Proteintech), p-SMAD2 ( Ab280888 , 1:200, Abcam), and HDAC6 (bs-2811R, 1:200, Bioss) were diluted appropriately and incubated with sections at 4 °C overnight.

Techniques: Ubiquitin Proteomics

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 levels were increased in the serum of patients with myocardial fibrosis, in the hearts of mice after CFPMI and in cardiac fibroblasts stimulated with fibrotic factors. A . ADAMTS1 levels in clinical serum samples were evaluated via ELISA. N = 30. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. B . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. C . Survival rate of mice in each group. D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E and F . ADAMTS1 expression in mouse heart tissue was determined through Western blot and IHC. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 12, 24, or 48 h to construct in vitro models. The cells were divided into four groups: the Control, TGF-β1/Ang II (12 h), TGF-β1/Ang II (24 h), and TGF-β1/Ang II (48 h) groups. G and H . Western blot detection of the expression of ADAMTS1, Collagen I and FN. I. The expression levels of ADAMTS1, Collagen I and FN were measured via qRT‒PCR. After the optimal time point (48 h) was determined, the mice were further divided into the control, TGF-β1 and Ang II groups. J . ADAMTS1 expression in cells was assessed by IF. Human cardiac fibroblasts were treated with 10 ng/mL TGF-β1 or 0.1 μM Ang II for 15, 30, 60, 90, and 120 min. K. qRT-PCR detection of the mRNA expression of ADAMTS1, Collagen I and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Construct, Staining, Expressing, Western Blot, In Vitro, Control, Quantitative RT-PCR

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: SMAD2 regulated ADAMTS1 expression in human and mouse cardiac fibroblasts induced by TGF-β1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 15, 30, 60, 90, and 120 min. A and B . Western blot analysis of the expression of SMAD2, p-SMAD2 and ADAMTS1. Human and mouse cardiac fibroblasts were treated with 10 ng/mL TGF-β1 for 48 h, and the cells were divided into Control and TGF-β1 groups. C . Human and mouse cardiac fibroblasts were induced with 10 ng/mL TGF-β1 for 12 h, 24 h, 48 h, and 72 h. qRT‒PCR was performed to detect the mRNA levels of ADAMTS1, Collagen I, and FN. D . IF staining was conducted to determine the expression of p-SMAD2 and ADAMTS1 at 48 h. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. E . p-SMAD2 expression in mouse heart tissue was measured by IHC. F . ChIP verification of the interaction between SMAD2 and ADAMTS1. Then, we interfered with SMAD2 expression. The groups were the si-NC, si-SMAD2-1, and si-SMAD2-2 groups. G . SMAD2 expression was assessed via qRT‒PCR. After the best si-SMAD2 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-SMAD2, and TGF-β1 + si-SMAD2 groups. H . Western blot detection of ADAMTS1, Collagen I, and FN expression in cells. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Expressing, Western Blot, Control, Staining, Construct

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Overexpression of ADAMTS1 enhanced the production of collagen fiber proteins in human and mouse cardiac fibroblasts induced by TGF-β1. We overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1 (48 h), oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. A . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. B-D . IF staining of Collagen I and FN expression. Additionally, we interfered with ADAMTS1; the cells were grouped as follows: si-NC, si-ADAMTS1-1, and si-ADAMTS1-2. E . ADAMTS1 expression was determined through qRT‒PCR. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC, TGF-β1 (48 h), si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. F and G . Western blot detection of ADAMTS1, Collagen I, FN, and α-SMA expression. H and I . IF staining of Collagen I and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Over Expression, Western Blot, Expressing, Staining

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: Knockdown of ADAMTS1 alleviated TGF-β1-induced fibrosis by downregulating HDAC6 protein expression. A CFPMI mouse model was constructed, and the mice were randomly divided into a Sham group and a CFPMI group, with 6 mice/group. A . HDAC6 expression in mouse heart tissue was determined through IHC. After the best si-ADAMTS1 was selected, the cells were further divided into the si-NC and si-ADAMTS1 groups. B . ADAMTS1 and HDAC6 expression was assessed via qRT‒PCR. C . Western blot analysis of ADAMTS1 and HDAC6 expression. Furthermore, we overexpressed ADAMTS1 and divided the cells into the oe-NC, TGF-β1, oe-ADAMTS1, and TGF-β1 + oe-ADAMTS1 groups. D . Western blot detection of HDAC6 expression. After the si-ADAMTS1 group was selected, the cells were further divided into the si-NC, TGF-β1, si-ADAMTS1, and TGF-β1 + si-ADAMTS1 groups. E. Western blot detection of HDAC6 expression. Moreover, cells were treated with the HDAC6 inhibitor ACY1215 (5 μM) for 48 h and then divided into the si-NC, si-ADAMTS1, si-NC + ACY1215, and si-ADAMTS1 + ACY1215 groups. F. Western blot detection of TGF-β1, Collagen I, FN, and ADAMTS1 expression. Subsequently, experiments were performed in human and mouse cardiac fibroblasts with ADAMTS1 overexpression followed by treatment with the HDAC6 inhibitor ACY1215. G. Western blot analysis was conducted to detect the expression levels of ADAMTS1 and HDAC6. H. Western blot analysis was performed to determine the expression levels of TGF-β1, Collagen I, and FN. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Knockdown, Expressing, Construct, Western Blot, Over Expression

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: ADAMTS1 interacted with HDAC6 during fibrosis. A . Co-IP verification of the interaction of ADAMTS1 with HDAC6, with ADAMTS1 as the bait protein. In the experiment, we first captured the ADAMTS1 protein using a specific antibody and then used Co-IP technology to detect its interaction with HDAC6. B . Co-IP verification of the interaction of HDAC6 with ADAMTS1, with HDAC6 as the bait protein. In the experiment, we captured the HDAC6 protein using a specific antibody and then used Co-IP technology to detect its interaction with ADAMTS1. We used 10 ng/mL TGF-β1 to induce human and mouse cardiac fibroblasts for 48 h, and the cells were further divided into control and TGF-β1 groups. C . IF staining was performed to evaluate the colocalization of ADAMTS1 and HDAC6 in TGF-β1-treated human and mouse cardiac fibroblasts. D . Changes in the level of ubiquitinated HDAC6 protein in human and mouse cardiac fibroblasts transfected with oe-ADAMTS1/si-ADAMTS1 or oe-NC/si-NC in the presence of 10 μM MG132. E. In si-ADAMTS1-treated human and mouse cardiac fibroblasts, MG132 (10 μM) was added to detect the ubiquitination levels of ADAMTS1 protein. F. Human and mouse cardiac fibroblasts were treated with TGF-β1 and subjected to SMAD2 knockdown to determine the ubiquitination levels of ADAMTS1 protein. N = 3. *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Co-Immunoprecipitation Assay, Control, Staining, Transfection, Ubiquitin Proteomics, Knockdown

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-ADAMTS1 treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, and CFPMI + sh-ADAMTS1 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. D . HDAC6 expression was determined through qRT‒PCR. E. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. F. Western blot detection of Collagen I, and FN expression. N = 3. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: shRNA, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: AAV-shRNA-HDAC6 transfection combined with ADAMTS1 inhibitor treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI. The mice were randomly divided into the Sham, CFPMI, CFPMI + anti-ADAMTS1, CFPMI + sh-HDAC6, and CFPMI + anti-ADAMTS1 + sh-HDAC6 groups. A . Echocardiographic results of different treatment groups, with measurements of ejection fraction (EF) and fractional shortening (FS). N = 6. B . Survival rate of mice in each group. C and D . HE and picrosirius red staining of collagen deposition in mouse cardiac tissues, with quantification of the Collagen Volume Fraction. E. HDAC6 expression was determined through qRT‒PCR. F. Western blot analysis of the expression of TGF-β1, SMAD2, and p-SMAD2 in mouse heart tissue. G. Western blot detection of Collagen I and FN expression. N = 3. ** P < 0.01, *** P < 0.001

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: shRNA, Transfection, Staining, Expressing, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Targeting ADAMTS1/HDAC6 alleviates TGF-β1/SMAD2-associated cardiac fibrosis in cardiac fibrosis post-myocardial infarction

doi: 10.1007/s10565-026-10159-2

Figure Lengend Snippet: TGF-β1/SMAD2 regulated ADAMTS1 by mediating CFPMI through HDAC6 ubiquitination

Article Snippet: Primary antibodies against VIM (10,366–1-AP, 1:200, Proteintech), HDAC6 (12,834–1-AP, 1:50, Proteintech), ADAMTS1 (12,749–1-AP, 1:50, Proteintech), FN ( AWA10327 , 1:50, Abiowell), Collagen I (ab138492, 1:200, Abcam), and p-SMAD2 (ab280888, 1:50, Abcam) were incubated at 4 °C overnight.

Techniques: Ubiquitin Proteomics

Journal: Cancers

Article Title: Activation of G Protein-Coupled Estrogen Receptor Induces p53 and ADAMTS1 to Inhibit Tumor Growth and Suppress Liver Cancer Metastasis

doi: 10.3390/cancers17162623

Figure Lengend Snippet: ADAMTS1 and GPER agonists inhibit the invasive ability of SK-Hep-1 cells. ( a ) Genes with RNA expression increased by more than 5.0-fold ( p < 0.0005) and protein expression increased by more than 1.6-fold ( p < 0.001). ( b ) ADAMTS1-treated SK-Hep-1 cell lysates obtained after 48 h. The expression of EMT-related proteins was measured through Western blotting. The amount of each protein was calculated compared to β-actin using ImageJ. Data are presented as mean ± standard deviation ( n = 3). ( c ) An invasion assay was performed with ADAMTS1-treated SK-Hep-1 cells after 48 h. Photos were observed via microscopy (10×). Graph representing the area of the invading cell calculated using ImageJ. Data are presented as mean ± standard deviation ( n = 3). * p < 0.01 (two-tailed unpaired t -test). ( d ) The protein expression detected in SK-Hep-1 cells treated with G1 (1 µM) and tamoxifen (TA) (17 µM). Cell lysates from G1 and TA-treated SK-Hep-1 cells were collected after 48 h. ( e ) The invasion was confirmed with G1- and G15-treated SK-Hep-1 cells for 48 h. Photos were observed via microscopy (10×). The graph shows the area of invaded cells calculated using ImageJ. Data are presented as mean ± standard deviation ( n = 3) ** p < 0.01 (one-way ANOVA with Dunnett’s post hoc test). The original images of the Western Blotting figures can be found in .

Article Snippet: The following primary antibodies were used: GPER (1:1000, Abcam, Cambridge, UK), p-p53 (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA), cyclin-dependent kinase (CDK) 1 and Cyclin B (1:1000, Cell Signaling, Danvers, MA, USA), extracellular-signal-regulated kinase (ERK) and pERK (1:1000, Invitrogen, Waltham, MA, USA), p53 (1:1000, Millipore), p21 (1:2000, Millipore), ADAMTS1 (1:2000, Rockville, Origene, MD, USA), E-cadherin (1:1000, Cell Signaling), proliferating cell nuclear antigen (PCNA; 1:2000, Cell Signaling), vimentin (1:1000, Cell Signaling), and β-actin (1:5000, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: RNA Expression, Expressing, Western Blot, Standard Deviation, Invasion Assay, Microscopy, Two Tailed Test

Journal: Cancers

Article Title: Activation of G Protein-Coupled Estrogen Receptor Induces p53 and ADAMTS1 to Inhibit Tumor Growth and Suppress Liver Cancer Metastasis

doi: 10.3390/cancers17162623

Figure Lengend Snippet: GPER agonist increases ADAMTS1 expression and inhibits metastasis. ( a ) The SK-Hep-1 cell-derived xenograft model and the control group received intraperitoneal administration of G1 at 10 mg/kg thrice weekly. Tumor size and liver metastases were compared between the G1 and control groups (arrow: metastatic lesion). Data are presented as mean ± standard deviation ( n = 5) * p < 0.05 (two-tailed unpaired t -test). ( b ) ADAMTS1, E-cadherin, Vimentin, and PCNA levels were measured in the transplanted subcutaneous tumor. The ratio of these protein levels to β-actin was calculated using ImageJ. Data are presented as mean ± standard deviation ( n = 6) * p < 0.05, ** p < 0.01 (two-tailed unpaired t -test). ( c ) Immunohistochemical staining for ADAMTS1 in the subcutaneously implanted tumor. The area of ADAMTS1 protein levels was calculated using ImageJ. Data are presented as mean ± standard deviation ( n = 3) * p < 0.05 (two-tailed unpaired t -test). ( d ) The overall survival rates of liver cancer patients were compared with GPER and ADAMTS1 expression levels in men and women. In the KM plotter database, the gene IDs of GPER and ADAMTS1 were 2852 and 9510, respectively. HR, hazard ratio. Data were analyzed using the optimal threshold as the cut-off. The original images of the Western Blotting figures can be found in .

Article Snippet: The following primary antibodies were used: GPER (1:1000, Abcam, Cambridge, UK), p-p53 (1:1000, Santa Cruz Biotechnology, Dallas, TX, USA), cyclin-dependent kinase (CDK) 1 and Cyclin B (1:1000, Cell Signaling, Danvers, MA, USA), extracellular-signal-regulated kinase (ERK) and pERK (1:1000, Invitrogen, Waltham, MA, USA), p53 (1:1000, Millipore), p21 (1:2000, Millipore), ADAMTS1 (1:2000, Rockville, Origene, MD, USA), E-cadherin (1:1000, Cell Signaling), proliferating cell nuclear antigen (PCNA; 1:2000, Cell Signaling), vimentin (1:1000, Cell Signaling), and β-actin (1:5000, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Expressing, Derivative Assay, Control, Standard Deviation, Two Tailed Test, Immunohistochemical staining, Staining, Western Blot