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Figure 5. Left atrial fibrosis in mouse models with shorter telomeres. A, Representative pictures of immunofluorescence staining with antibody against collagen I and Titin and counterstaining with DAPI. B, Quantification of collagen I (fibrosis) in wild-type (WT) and Tert−/− fourth generation mice, n=5 per group. C, Gene expression of profibrotic and fibrotic markers in left atrium of WT and Tert−/− third generation mice. n=3–4 per group, data are expressed as mean±SEM. *P<0.05. Scale bars=50 μmol. Acta2 indicates Alpha-smooth muscle Actin; Fn, fibronectin; Ctgf, connective tissue growth factor; and <t>Col1A2,</t> <t>collagen</t> <t>type</t> <t>I</t> alpha 2 chain.
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Effect of miR-483-3p by qPCR Analysis. HTM cells were transfected with hsa- miR-483-3p mimic, mimic negative control and inhibitor after 100nM DEX treatment. Total RNA was extracted, converted to cDNA and <t>the</t> <t>SMAD4,TGFβ2,Collagen</t> <t>1A,</t> Fibronectin and Laminin5 gene expression were carried out by qPCR, gene expression were normalized to ACTB and analyzed using the 2 −ΔΔCT method. SMAD4- GC-R (a) GC-NR (b) HTM cells (n=3); TGFβ2 - GC-R (c) GC-NR (d) HTM cells (n=3); Collagen 1A GC-R (e) GC-NR (f) HTM cells (n=3); Fibronectin GC-R (g) GC-NR (h) HTM cells (n=3); Laminin5 GC-R (i) GC-NR (j) HTM cells (n=3). Data represent mean ± SEM; *p <0.05. **p <0.001; ***p <0.0001.
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Image Search Results


Figure 5. Left atrial fibrosis in mouse models with shorter telomeres. A, Representative pictures of immunofluorescence staining with antibody against collagen I and Titin and counterstaining with DAPI. B, Quantification of collagen I (fibrosis) in wild-type (WT) and Tert−/− fourth generation mice, n=5 per group. C, Gene expression of profibrotic and fibrotic markers in left atrium of WT and Tert−/− third generation mice. n=3–4 per group, data are expressed as mean±SEM. *P<0.05. Scale bars=50 μmol. Acta2 indicates Alpha-smooth muscle Actin; Fn, fibronectin; Ctgf, connective tissue growth factor; and Col1A2, collagen type I alpha 2 chain.

Journal: Journal of the American Heart Association

Article Title: Telomere Length Is Associated With Adverse Atrial Remodeling in Patients With Atrial Fibrillation

doi: 10.1161/jaha.124.037512

Figure Lengend Snippet: Figure 5. Left atrial fibrosis in mouse models with shorter telomeres. A, Representative pictures of immunofluorescence staining with antibody against collagen I and Titin and counterstaining with DAPI. B, Quantification of collagen I (fibrosis) in wild-type (WT) and Tert−/− fourth generation mice, n=5 per group. C, Gene expression of profibrotic and fibrotic markers in left atrium of WT and Tert−/− third generation mice. n=3–4 per group, data are expressed as mean±SEM. *P<0.05. Scale bars=50 μmol. Acta2 indicates Alpha-smooth muscle Actin; Fn, fibronectin; Ctgf, connective tissue growth factor; and Col1A2, collagen type I alpha 2 chain.

Article Snippet: Real time PCR quantification of fibrosis markers smooth muscle actin (Acta2), fibronectin (Fn), connective tissue growth factor (Ctgf) and collagen 1a (Col1A2) was done using the Takyon sybr green (Eurogentec) on the StepOnePlus system (Applied Biosystems).

Techniques: Immunofluorescence, Staining, Gene Expression

Effect of miR-483-3p by qPCR Analysis. HTM cells were transfected with hsa- miR-483-3p mimic, mimic negative control and inhibitor after 100nM DEX treatment. Total RNA was extracted, converted to cDNA and the SMAD4,TGFβ2,Collagen 1A, Fibronectin and Laminin5 gene expression were carried out by qPCR, gene expression were normalized to ACTB and analyzed using the 2 −ΔΔCT method. SMAD4- GC-R (a) GC-NR (b) HTM cells (n=3); TGFβ2 - GC-R (c) GC-NR (d) HTM cells (n=3); Collagen 1A GC-R (e) GC-NR (f) HTM cells (n=3); Fibronectin GC-R (g) GC-NR (h) HTM cells (n=3); Laminin5 GC-R (i) GC-NR (j) HTM cells (n=3). Data represent mean ± SEM; *p <0.05. **p <0.001; ***p <0.0001.

Journal: bioRxiv

Article Title: Hsa-MiR-483 -3p Regulates the Extracellular Matrix Proteins via TGFβ2/SMAD4 Signaling in the Glucocorticoid-responsive Human Trabecular Meshwork Cells

doi: 10.1101/2025.01.14.633091

Figure Lengend Snippet: Effect of miR-483-3p by qPCR Analysis. HTM cells were transfected with hsa- miR-483-3p mimic, mimic negative control and inhibitor after 100nM DEX treatment. Total RNA was extracted, converted to cDNA and the SMAD4,TGFβ2,Collagen 1A, Fibronectin and Laminin5 gene expression were carried out by qPCR, gene expression were normalized to ACTB and analyzed using the 2 −ΔΔCT method. SMAD4- GC-R (a) GC-NR (b) HTM cells (n=3); TGFβ2 - GC-R (c) GC-NR (d) HTM cells (n=3); Collagen 1A GC-R (e) GC-NR (f) HTM cells (n=3); Fibronectin GC-R (g) GC-NR (h) HTM cells (n=3); Laminin5 GC-R (i) GC-NR (j) HTM cells (n=3). Data represent mean ± SEM; *p <0.05. **p <0.001; ***p <0.0001.

Article Snippet: Membrane was blocked with 5% nonfat dry milk and incubated with primary antibodies [SMAD4 (1:1000), Abcam (Cat# ab110175); TGFβ2 (1:500), Abcam (Cat# ab167655); collagen 1A (1:250), Santa Cruz (Cat# sc-59772); fibronectin (1:100), Santa Cruz (Cat# sc-52331), laminin5 (1:500), Santa Cruz (Cat# sc-13587)] overnight at 4°C.

Techniques: Transfection, Negative Control, Expressing

Proposed Schematic Representation of the Effect of miR483-3p on ECM Regulation in GC-R HTM cells. In GC-R HTM cells treatment with dexamethasone (DEX) activates canonical TGFβ2-SMAD signaling and causes dysregulated extracellular matrix (ECM) protein turnover resulting in the accumulation of ECM proteins which is implicated in steroid-induced ocular hypertension in GC-R HTM cells. The upregulation of TGFβ2-SMAD signaling by DEX (Panel A) in HTM cells. The upregulation of hsa-miR 483-3p inhibits the activation of TGFβ2-SMAD signaling by binding to SMAD4 mRNA leading to decreased SMAD4 protein (Panel B) in HTM cells. However, the presence of hsa-miR 483-3p antagomir/inhibitor decreases the expression of miR483-3p which in-turn enhances ECM deposition through the activation of TGFβ2-SMAD signaling (Panel C). Interestingly, the regulation of ECM by has-miR 483-3p was more pronounced in DEX-responder HTM cells (Left) as compared to DEX-non-responder cells (Right). TGFβR –TGFβ receptor complex; Col1A – Collagen 1A; FN – Fibronectin; CTGF-Connective tissue growth factor.

Journal: bioRxiv

Article Title: Hsa-MiR-483 -3p Regulates the Extracellular Matrix Proteins via TGFβ2/SMAD4 Signaling in the Glucocorticoid-responsive Human Trabecular Meshwork Cells

doi: 10.1101/2025.01.14.633091

Figure Lengend Snippet: Proposed Schematic Representation of the Effect of miR483-3p on ECM Regulation in GC-R HTM cells. In GC-R HTM cells treatment with dexamethasone (DEX) activates canonical TGFβ2-SMAD signaling and causes dysregulated extracellular matrix (ECM) protein turnover resulting in the accumulation of ECM proteins which is implicated in steroid-induced ocular hypertension in GC-R HTM cells. The upregulation of TGFβ2-SMAD signaling by DEX (Panel A) in HTM cells. The upregulation of hsa-miR 483-3p inhibits the activation of TGFβ2-SMAD signaling by binding to SMAD4 mRNA leading to decreased SMAD4 protein (Panel B) in HTM cells. However, the presence of hsa-miR 483-3p antagomir/inhibitor decreases the expression of miR483-3p which in-turn enhances ECM deposition through the activation of TGFβ2-SMAD signaling (Panel C). Interestingly, the regulation of ECM by has-miR 483-3p was more pronounced in DEX-responder HTM cells (Left) as compared to DEX-non-responder cells (Right). TGFβR –TGFβ receptor complex; Col1A – Collagen 1A; FN – Fibronectin; CTGF-Connective tissue growth factor.

Article Snippet: Membrane was blocked with 5% nonfat dry milk and incubated with primary antibodies [SMAD4 (1:1000), Abcam (Cat# ab110175); TGFβ2 (1:500), Abcam (Cat# ab167655); collagen 1A (1:250), Santa Cruz (Cat# sc-59772); fibronectin (1:100), Santa Cruz (Cat# sc-52331), laminin5 (1:500), Santa Cruz (Cat# sc-13587)] overnight at 4°C.

Techniques: Activation Assay, Binding Assay, Expressing