Journal: Molecular therapy. Nucleic acids

Article Title: Angiotensin II mediates hypertensive cardiac fibrosis via an Erbb4-IR-dependent mechanism.

doi: 10.1016/j.omtn.2023.06.017

Figure Lengend Snippet: Figure 2. Silencing cardiac Erbb4-IR prevents Ang II-induced cardiac fibrosis in hypertensive mice (A) Representative images of H&E and Masson’s tri- chrome staining. (B) Representative immunohistochem- ical images of collagen I, collagen III, a-SMA, and fibro- nectin. (C) Quantitative analysis of collagen I, collagen III, a-SMA, and fibronectin from immunohistochemical staining. Data are presented as mean ± SEM for a group of eight mice. **p < 0.01, ***p < 0.001 compared with SL; #p < 0.05, ##p < 0.01 compared with Ang II + EV control. Scale bars, 20 mm.

Article Snippet: Total protein from the LV was extracted for western blot analysis as described previously.10,11,24–26 In brief, after blocking nonspecific binding with 5% BSA, the membrane was incubated at 4 C overnight with the indicated primary antibodies, including those against collagen I and III (catalog numbers 1310-01 and 1330-01, respectively; Southern Biotech), a-SMA (catalog number A5228, Sigma), fibronectin (catalog number sc-8422, Santa Cruz Biotechnology), p-Smad2 at the pS465 and pS467 residues/Smad3 at the pS423 and pS425 residues (catalog number 9510, Cell Signaling Technology, USA), Smad2/3 (catalog number 3102, Cell Signaling Technology), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Chemicon, Temecula, CA, USA).

Techniques: Staining, Immunohistochemical staining, Control

Journal: Molecular therapy. Nucleic acids

Article Title: Angiotensin II mediates hypertensive cardiac fibrosis via an Erbb4-IR-dependent mechanism.

doi: 10.1016/j.omtn.2023.06.017

Figure Lengend Snippet: Figure 7. In vitro studies detect that addition of Ang II induces Erbb4-IR expression by CFs but not by cardiomyocytes and that silencing Erbb4-IR protects against Ang II-induced extracellular matrix production by CFs (A) Addition of Ang II (1 mM) for 12 h markedly upregulates Erbb4-IR in CFs but not in cardiomyocytes. (B) Addition of Ang II (1 mM) upregulates Erbb4-IR in CFs in a time- dependent manner. (C) Expression of Erbb4-IR by CFs treated with or without Ang II (1 mM) and/or Erbb4-IR shRNA. (D–G) Real-time PCR reveals that silencing Erbb4-IR inhibits Ang II (1 mM)-induced mRNA expression of a-SMA, collagen I, collagen III, and fibronectin by CFs. (H–L) Western blot analysis shows that silencing Erbb4-IR inhibits Ang II (1 mM)-induced protein expression of a-SMA, collagen I, collagen III, and fibronectin by CFs. Data are the mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 compared with negative control (Ctrl) without Ang II treatment; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with Ang II + negative EV Ctrl (NC).

Article Snippet: Total protein from the LV was extracted for western blot analysis as described previously.10,11,24–26 In brief, after blocking nonspecific binding with 5% BSA, the membrane was incubated at 4 C overnight with the indicated primary antibodies, including those against collagen I and III (catalog numbers 1310-01 and 1330-01, respectively; Southern Biotech), a-SMA (catalog number A5228, Sigma), fibronectin (catalog number sc-8422, Santa Cruz Biotechnology), p-Smad2 at the pS465 and pS467 residues/Smad3 at the pS423 and pS425 residues (catalog number 9510, Cell Signaling Technology, USA), Smad2/3 (catalog number 3102, Cell Signaling Technology), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Chemicon, Temecula, CA, USA).

Techniques: In Vitro, Expressing, shRNA, Real-time Polymerase Chain Reaction, Western Blot, Negative Control