col3a antibody  (SouthernBiotech)

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet: DCM-Exos induced up-regulation of fibrotic genes in CFs (A) Representative fluorescent images of CTL-Exos or DCM-Exos (PKH26-labeled) treatment in CFs. Endocytosed Exos (orange) were observed in the cytoplasm of CFs (bar = 200 μm (left); 900 nm (right)). (B) Quantifications of Exo uptake in CFs. The uptake ratio was evaluated by the number of Exo uptaken cells (orange) divided by the total number of nuclei (blue) in the image (N = 22 pics/group), NS, nonsignificant. (C) qRT-PCR of fibrotic genes in CFs. N = 6; ∗ = p< 0.05. (D) Representative immunoblots. The CF cell lysis of different treatment groups was probed with specific antibodies (Col1a, Col3a, and CTGF). (E) Quantification of protein expression in immunoblots. Col1a, col3a, and CTGF expression were normalized to the loading control (GAPDH), N = 6. Data represented as mean ± SD; ∗ = p< 0.05; one-way ANOVA followed by Tukey post hoc test.

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

Techniques: Labeling, Quantitative RT-PCR, Western Blot, Lysis, Expressing, Control

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet: Intracardial injection of DCM-Exo promoted fibrosis in mouse hearts (A) Representative echocardiographic images. Images were taken respectively from PBS, CTL-Exo, or DCM-Exo injection groups on day 0 and day 14. (B) LVEF was determined by echocardiography. The injection of DCM-Exo significantly decreased LVEF on day 14 (N = 8/group; ∗ = p< 0.05). Injection of PBS, or CTL-Exo resulted in nonsignificant change between day 14 and baseline. (C) Representative pictures of picrosirius red/fast green staining in series sections of the mouse hearts. The fibrotic area (Red) was quantified by ImageJ. N = 8/group, ∗∗ = p< 0.01. (D) Representative immunoblots of mouse hearts (PBS, CTL-Exo, or DCM-Exo treatments). Fibrotic markers, Col1a, Col3a, and CTGF were detected by specific antibodies. (E) Quantification of fibrotic markers expression. DCM-Exos injection resulted in significant upregulation of fibrotic markers (N= 8/group). GAPDH was used as a protein loading control. Data represented as mean ± SD; ∗ = p< 0.05; one-way ANOVA followed by Tukey post hoc test.

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

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

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet: MiR-218-5p increased Smad2 phosphorylation in CFs (A) Immunoblots of cell lysates derived from CFs treated with TGF-β, miR-218-5p, and a TGF-β inhibitor (SB525334). Specific antibodies were used to detect Col1a, Col3a, CTGF, phospo-Smad2 ( p -SMAD2), and GAPDH. (B) Quantitation of protein expression in immunoblots. GAPDH was used as a protein loading control. TGF-β inhibitor (SB525334) significantly reduced TGF-β1-induced fibrotic effect (lane 3). MiR-218-5p increased the phosphorylation of Smad2. N= 3; one-way ANONVA followed by Tukey’s post hoc test, ∗ = p< 0.05; ∗∗ = p< 0.01; ∗∗∗ = p< 0.001. Data represented as mean ± SD.

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

Techniques: Phospho-proteomics, Western Blot, Derivative Assay, Quantitation Assay, Expressing, Control

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet: MiR-218-5p targeted to TNFAIP3 (A) Targeted sequence prediction of miR-218-5p in RNA22. (B) Immunoblots (left) indicated TNFAIP3 expression was significantly repressed in CFs transfected with miR-218-5p. (C) Luciferase assay of miR-218-5p and TNFNAIP3 5′UTR region. Co-transfection of miR-218-5p + WT sequence significantly reduced luciferase activity compared with the scramble CTL + WT. One-way ANOVA followed by Tukey’s post hoc test, ∗∗ = p< 0.01; NS = nonsignificant. (D) Representative immunoblots of CFs treatment. (E) Quantitation of expression of TNFAIP3, Col1a, Col3a, and CTGF. One-way ANOVA followed by Tukey’s post hoc test; ∗∗ = p<0.01, ∗∗∗ = p< 0.001. Data represented as mean ± SD.

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

Techniques: Sequencing, Western Blot, Expressing, Transfection, Luciferase, Cotransfection, Activity Assay, Quantitation Assay

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet: Restoration of TNFAIP3 ameliorated the DCM-Exo-induced fibrotic effect (A) Representative immunoblots of CFs treated with either CTL-Exos or DCM-Exos. DCM-Exo treatment significantly decreased TNFAIP3 expression. Student’s t test; ∗∗ = p <0.01. (B) Representative immunoblots of TNFAIP3, Col1a, Col3a, CTGF, and GAPDH detection in CFs treated with CTL-Exos, DCM-Exos, or overexpression of TNFAIP3. (C–F) Quantitation of protein expression. TNFAIP3 overexpression significantly reduced the expression of fibrotic markers. One-ANOVA followed by Tukey’s post hoc test; N= 3; ∗∗ = p< 0.01. (G) Schematic picture showing the working model of our study. Data represented as mean ± SD.

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

Techniques: Western Blot, Expressing, Over Expression, Quantitation Assay

Journal: iScience

Article Title: Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway

doi: 10.1016/j.isci.2023.105963

Figure Lengend Snippet:

Article Snippet: Col3a , Cell Signaling Technology (H) , 66887.

Techniques: Virus, Recombinant, Mutagenesis, Software

Journal: iScience

Article Title: Bizonal cardiac engineered tissues with differential maturation features in a mid-throughput multimodal bioreactor

doi: 10.1016/j.isci.2022.104297

Figure Lengend Snippet:

Article Snippet: Collagen type III Col3a , Thermo-Fisher Scientific , Rn01437681_m1.

Techniques: Recombinant, Electron Microscopy, Bicinchoninic Acid Protein Assay, Staining, Software, Modification, Membrane, Saline