Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 3. TXNIP promoted post-MI collagen deposition (A,B) Representative picrosirius red staining images of heart sections from each group at 28 days after treatment (A) under a light microscope and (B) under a polarized light microscope. Collagen I showed red/orange birefringence, while Collagen III showed green/whitish birefringence. (C,E,G) Representative immunohistochemical staining images of heart sections from each group at 28 days after treatment. (C) Collagen I deposition. (E) Collagen III deposition, and (G) ACTA2 deposition. (D) Statistical analysis of Collagen I deposition. Data are shown as the mean±SEM, n=3. **P<0.01. (F) Statistical analysis of Collagen III deposition. Data are shown as the mean± SEM, n=3. **P<0.01. (H) Statistical analysis of ACTA2 deposition. Data are shown as the mean±SEM, n=3. *P<0.05, **P<0.01.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques: Staining, Light Microscopy, Immunohistochemical staining

Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 4. TXNIP increased post-MI collagen deposition (A) Representative images of the protein levels of Collagen III, Collagen I, ACTA2, and CTGF. (B‒E) Statistical analyses of the protein levels of Collagen I, Collagen III, ACTA2, and CTGF. Data are shown as the mean±SEM, n=3‒5. *P<0.05, **P<0.01; ns, not significant.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques:

Journal: The Journal of pathology

Article Title: A 3D tri-culture system reveals that activin receptor-like kinase 5 and connective tissue growth factor drive human glomerulosclerosis

doi: 10.1002/path.4960

Figure Lengend Snippet: (A) MC 3D monoculture without the addition of TGFβ, and (B) in the presence of TGFβ, shown by H&E staining of paraffin-embedded sections. Human collagen type I/III (immunostained with an antibody that recognises both collagen type I and III) (C) and human collagen type IV (D) can be demonstrated in nodules by immunoperoxidase staining. (E) High-power view of MC nodule by scanning electron microscopy in cross section. (F) MC nodule count and collagen type I alpha1 (COL1a1), and collagen type IV alpha1 (COL4a1) RNA quantification (n=3). (G) Effect of ALK5 inhibition (Alk5i) on MC nodule formation (n=3). (H) Effect of SMAD2 or SMAD3 siRNA knockdown in MCs on nodule formation (n=3) with immunoblots demonstrating degree of knockdown. [DharmaFECT (DH), non-targeting siRNA control (siCP), siSMAD2 (siS2), siSMAD3 (siS3), total SMAD2 (tSMAD2), total SMAD3 (tSMAD3)]. Error bars represent 1 s.d. NS, not significant; ***, p<0.001; **, p<0.01: 2-tailed Student’s t test.

Article Snippet: After culture, matrices were embedded in HistoGel, fixed in 4% paraformaldehyde and paraffin wax-embedded for sectioning and staining for human collagen type IV (mouse anti-human collagen IV monoclonal antibody 2150–0121, Bio-Rad, CA, USA) and human collagen type I/III (rabbit anti-human collagen I/III polyclonal antibody 2150–2210, Bio-Rad, CA, USA).

Techniques: Staining, Immunoperoxidase Staining, Electron Microscopy, Inhibition, Western Blot

Journal: Scientific Reports

Article Title: Exercise reduces hyperlipidemia-induced cardiac damage in apolipoprotein E-deficient mice via its effects against inflammation and oxidative stress

doi: 10.1038/s41598-023-36145-w

Figure Lengend Snippet: Effect of exercise on hyperlipidemia-induced cardiac fibrosis. ( A ) Fibrosis was evaluated using Masson’s trichrome staining. Scale bar = 100 μm. Arrows indicate positively stained cells. ( B ) Representative immunohistochemistry staining for TGF-β, collagen III, and Smad3. Scale bar = 100 μm. Arrows indicate positively stained cells. ( C ) Bar graph showing Masson’s trichrome, TGF-β, collagen III and Smad3 positive cells. ( D ) Western blotting for TGF-β, collagen III, and Smad3 protein expression in cardiac tissue. ( E ) Quantification of TGF-β, collagen III, and Smad3 protein expression. Data represent the mean ± SEM; n = 3 per group. *P < 0.05. ND normal diet, E exercise training, HFD high-fat diet, ApoE apolipoprotein E, TGF-β transforming growth factor-β.

Article Snippet: The sections were treated for 15 min with 3% H 2 O 2 in methanol to inactivate endogenous peroxidases and then incubated at 4 °C overnight with primary antibodies to SIRT1 (rabbit anti-SIRT1 antibody, 1:300; Solarbio), NRF2 (rabbit anti-NRF2 antibody, 1:200; Proteintech), NOX4 (rabbit anti-NOX4 antibody, 1:200; Proteintech), TGF-β (rabbit anti-TGF-β antibody, 1:200; Solarbio), HO-1 (rabbit anti-HO-1 antibody, 1:100; Solarbio), collagen III (rabbit anti-collagen III antibody, 1:100; Solarbio), Smad3 (rabbit anti-Smad3 antibody, 1:100; Solarbio), Bax (rabbit anti-Bax antibody, 1:50; Solarbio), Bak (rabbit anti-Bak antibody, 1:100; Solarbio), Bcl-2 (rabbit anti-Bcl-2 antibody, 1:50; Solarbio) and Bcl-xl (rabbit anti-Bcl-xl antibody, 1:100; Solarbio).

Techniques: Staining, Immunohistochemistry, Western Blot, Expressing