mmp-2 inhibitor i  (Cayman Chemical)

Journal: Biomolecules & biomedicine

Article Title: SKIL/SnoN attenuates TGF-β1/SMAD signaling-dependent collagen synthesis in hepatic fibrosis.

doi: 10.17305/bb.2023.9000

Figure Lengend Snippet: Figure 3. SnoN regulates fibrosis-associated proteins through its interaction with p-SMAD2 and p-SMAD3. (A) Gross specimen morphology for the BDL and control groups. Histologic examination of liver sections was performed using hematoxylin and eosin and Masson’s trichrome staining (magnification × 200); (B) The positive expression location of TGF-β1, TIMP-1, collagen III, and SnoN analyzed by immunohistochemistry assay (magnification × 200); (C) The expressions of TGF-β1, collagen III, and SnoN protein in liver tissue of rats in each group were observed by immunohistochemistry, the relative protein expression of SnoN in liver tissue of rats in each group was detected by western blot. * P < 0.05 compared with the control group; (D) Quantitative PCR analysis of the mRNA expression of TGF-β1, TIMP-1, collagen III, collagen I, MMP-13, and SnoN; (E) As time went on, the expression of TGF-β1, TIMP-1, collagen III, and FN proteins increased, while SnoN protein decreased. Data are expressed as mean ± SD, n = 10 rats/group. * P < 0.05 compared with the control group; (F) Immunofluorescent staining showed the co-location of SnoN and p-SMAD2, the co-location of SnoN and p-SMAD3; (G) The interaction of SnoN and p-SMAD2 and the interaction of SnoN and p-SMAD3 in HSC-T6 cells were examined by co-IP, followed by immunoblot analysis. IgG represents a control antibody used for IPs. BDL: Bile duct ligation; HE: Hematoxylin and eosin; TGF-β1: Transforming growth factor beta 1; TIMP-1: Tissue inhibitor of metalloproteinase 1; SnoN: Ski-related novel protein N; MMP-13: Matrix metalloproteinase 13; FN: Fibronectin; SMAD: Suppressor of mothers against decapentaplegic protein; HSC: Hepatic stellate cells; IP: Immunoprecipitation; co-IP: Co-immunoprecipitation.

Article Snippet: The following primary antibodies were used in immunohistochemistry: polyclonal rabbit anti-rat SnoN (1:200 dilution; Abcam, Cambridge, UK), collagen III (1:50 dilution, Abcam), TGF-β1 (1:50 dilution; Santa Cruz, OR, USA), tissue inhibitor of metalloproteinase 1 (TIMP-1) (1:50 dilution, Santa Cruz), and matrix metalloproteinase 13 (MMP-13) (1:50 dilution, Santa Cruz).

Techniques: Control, Staining, Expressing, Immunohistochemistry, Western Blot, Real-time Polymerase Chain Reaction, Co-Immunoprecipitation Assay, Ligation, Immunoprecipitation

Journal: Biomolecules & biomedicine

Article Title: SKIL/SnoN attenuates TGF-β1/SMAD signaling-dependent collagen synthesis in hepatic fibrosis.

doi: 10.17305/bb.2023.9000

Figure Lengend Snippet: Figure 4. Effect of SnoN on apoptosis and fibrosis-associated proteins. (A) The expression of SnoN protein after transfection. *P < 0.05 compared with the control group; (B) Apoptosis of cells in each group was detected by flow cytometry with Annexin V-FITC/PI labeling. *P < 0.05 compared with the control group; (C) Western blot analysis of protein expression of collagen I, collagen III, TIMP-1, and MMP-13 after transfection. *P < 0.05 compared with the control group; #P < 0.05 compared with the Ad-EGFP group; %P < 0.05 compared with the si-FITC group; (D) Western blot analysis of protein expression of collagen I. SnoN: Ski-related novel protein N; EGFP: Enhanced green fluorescent protein; FITC: Fluorescein isothiocyanate; PI: Propidium iodide; TIMP-1: Tissue inhibitor of metalloproteinase 1; MMP-13: Matrix metalloproteinase-13; TGF-β1: Transforming growth factor beta 1.

Article Snippet: The following primary antibodies were used in immunohistochemistry: polyclonal rabbit anti-rat SnoN (1:200 dilution; Abcam, Cambridge, UK), collagen III (1:50 dilution, Abcam), TGF-β1 (1:50 dilution; Santa Cruz, OR, USA), tissue inhibitor of metalloproteinase 1 (TIMP-1) (1:50 dilution, Santa Cruz), and matrix metalloproteinase 13 (MMP-13) (1:50 dilution, Santa Cruz).

Techniques: Expressing, Transfection, Control, Flow Cytometry, Labeling, Western Blot

Journal: Mediators of inflammation

Article Title: Empagliflozin Reverses Oxidized LDL-Induced RECK Suppression, Cardiotrophin-1 Expression, MMP Activation, and Human Aortic Smooth Muscle Cell Proliferation and Migration.

doi: 10.1155/2023/6112301

Figure Lengend Snippet: FIGURE 2: OxLDL stimulates SMC migration via MMP2 and MMP9 activation. (a, b) OxLDL stimulates MMP2 and MMP9 activation. SMC made quiescent in medium containing ITS-G 1x supplement were exposed to OxLDL or nLDL for 24 hr and then analyzed for MMP2 and

Article Snippet: AR-100, a biphenylsulfonamide and selective inhibitor of MMP2, and AG-L-66085, a cell-permeable and reversible MMP9 inhibitor, were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX).

Techniques: Migration, Activation Assay

Journal: JACC: Basic to Translational Science

Article Title: Beta 1 -Adrenergic Receptor Cleavage and Regulation by Elastase

doi: 10.1016/j.jacbts.2023.02.002

Figure Lengend Snippet: Elastase Cleaves the Full-Length β 1 AR in CHO-Lec2 Cells at a Site That Is Distinct From Previously Identified Cleavage Sites at R 31 ↓L 32 , S 41 ↓L 42 , or P 52 ↓L 53 Lysates from WT-β 1 AR- or β 1 AR-31/32-expressing CHO-Pro5 treated with vehicle or GM6001 (10 μM for 24 hours; A ) or from β 1 AR-31/32, β 1 AR-41/42, or β 1 AR-52/53 cleavage-resistant mutant expressing CHO-Lec2 cells treated with vehicle or elastase (20 μg/mL for 10 minutes; B ) were probed for anti-β 1 AR immunoreactivity. A representative experiment is depicted on top, with results from 5 (A) or 4 (B) separate experiments performed on separate culture preparations quantified at the bottom. Data are shown as mean ± SEM, with the percent of full-length β 1 AR (upper band) expressed as a % of total (full-length + truncated) β 1 AR (∗ P < 0.05). (C) Immunoblot analysis of lysates from β 1 AR-31/41/52 expressing CHO-Lec2 cells cultured in the presence of vehicle, MMP 2/3 inhibitor (20 μM), MMP 2/9 inhibitor (10 μM), MMP 9/13 inhibitor (20 nM), ADAM 10 inhibitor (400 nM), ADAM10/17 inhibitor (3 μM), TAPI-2 (20 μM), or GM6001 (10 μM) for 24 hours. Results were replicated in 2 separate experiments. Abbreviations as in Figure 1 .

Article Snippet: Elastase from porcine pancreas (Cat# E0258), GM6001 (Cat# 364206), GI254023X (ADAM-10 inhibitor, Cat# SML0789), matrix metalloproteinase (MMP)-2/MMP-9 inhibitor I (Cat# 444241), MMP-9/MMP-13 inhibitor I (Cat# 444252), forskolin (Cat# F3917), isoproterenol (Iso) (Cat# 420355), propranolol (Cat# P0884), sotalol (Cat# S0278), and theophylline (Cat# T1633) were from Sigma-Aldrich.

Techniques: Expressing, Mutagenesis, Western Blot, Cell Culture