Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 1. Rats of diabetic cardiomyopathy (DCM) group showed insulin resistance after four-week HF diet and ALK7 silencing ameliorated insulin resistance. (a, c) Intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT) were performed at baseline and at Week 4 in rats of DCM group. (b, d) Area under the receiver operating characteristic curve (AUC) for IPGTT and IPITT. (E, G) IPGTT and IPITT were repeated at the end of the experiment for all groups. (f, h) AUC for IPGTT and IPITT. Data are mean ± SD; n = 7–10 per group. HF: high fat. *P < 0.05; **P < 0.01; ***P < 0.001 vs. baseline; ‡P < 0.001 vs. control; #P < 0.05; ##P < 0.01; ###P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 2. ALK7 gene silencing improved metabolic disturbance. (A) Fasting insulin (FINS). (B) Insulin sensitivity index (ISI). (C) Total cholesterol (TC) levels. (D) Triglyceride (TG) levels. (E) Fasting blood glucose (FBG). Data are mean ± SD; n = 7–10 per group. *P < 0.05; ***P < 0.001 vs. control; ††P < 0.01; ‡P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 3. ALK7 silencing ameliorated left ventricular (LV) dysfunction in diabetic rats. (A) Pressure curves of cardiac catheterization. (B) Analysis of LV end-diastolic pressure (LVEDP) LV systolic pressure (LVSP) in all groups. Data are mean ± SD; n = 7–8 per group. (A color version of this figure is available in the online journal.) *P < 0.05; **P < 0.01; ***P < 0.001 vs. control; ‡P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 4. ALK7 gene silencing ameliorated cardiac remodeling in DCM. (Aa). Heart size (scale bar: 5 mm). (Ab). Representative LV cross-section stained with hematoxylin and eosin (H&E) at papillary muscle level (scale bar: 5 mm). (Ac). Longitudinal section of LV stained with H&E (scale bar: 50 μm). (Ad). Transverse sections of LV stained with H&E (scale bar: 100 μm). (B). The heart weight-to-body weight ratio (HW/BW). (C) Quantitative analysis of LV cardiomyocyte size. Data are mean ± SD; n = 7–10 per group. **P < 0.01; ***P < 0.001 vs. control; †P < 0.05; ‡P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Staining, Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 5. ALK7 silencing attenuated cardiomyocyte apoptosis in diabetic rats. (A) Representatives of TUNEL staining in cardiomyocytes from LV tissues (brown staining considered TUNEL-positive; scale bar: 100 μm). (B) Quantitative analysis of TUNEL-positive cardiomyocytes (n = 6 per group). (C) Representative western blot of Bax, Bcl2, and cleaved Caspase3. (D) Western blot analysis of Bcl2-to-Bax ratio (Bcl2/Bax). (E) Western blot analysis of cleaved Caspase3. Values are the mean ± SD, and western blots were conducted at least 3 times. (A color version of this figure is available in the online journal.) *P < 0.05; ***P < 0.001 vs. control; ††P < 0.01; ‡P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: TUNEL Assay, Staining, Western Blot, Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 6. ALK7 gene silencing attenuated cardiac fibrosis in diabetic rats. Aa to Ac: Interstitial fibrosis. (Aa) Representative Masson trichrome staining (collagen fibers is stained green and myocardium is red). (Ab, Ac) Sirius red staining (collagen is stained bright red; Ab: bright-field, Ac: dark-field). Ad to Af: Perivascular fibrosis. (Ad) Masson trichrome staining. (Ae) Sirius red staining (bright-field). (Af) Sirius red staining (dark-field). (B) Quantitative analysis of CVF. (C) Quantitative analysis of PVCA/LA. (D) Representative immunohistochemical staining of Collagen I (Da) and III (Db). (E) Quantitative analysis of Collagen I-to-III ratio (Coll-I/Coll-III). (F) Representative western blot of Coll-I and Coll-III. (G) Western blot analysis of Coll-I. (H) Western blot analysis of Coll-III. (I) Western blot analysis of Coll-I/Coll-III. Scale bar: 50 μm; Data are mean ± SD; n = 6 per group. *P < 0.05; **P < 0.01; ***P < 0.001 vs. control; ††P < 0.01; ‡P < 0.001 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Staining, Immunohistochemical staining, Western Blot, Control

Journal: Experimental biology and medicine (Maywood, N.J.)

Article Title: Activin receptor-like kinase 7 silencing alleviates cardiomyocyte apoptosis, cardiac fibrosis, and dysfunction in diabetic rats.

doi: 10.1177/15353702221095049

Figure Lengend Snippet: Figure 7. ALK7 silencing restored Akt phosphorylation and suppressed Smad2/3 phosphorylation. (A) Relative ALK7 mRNA expression in all groups. (B) The silencing efficacy of ALK7-siRNA measured by western blot. (C) Representative western blot of p-Akt and t-Akt. (D) Western blot analysis of p-Akt/t-Akt. (E) Representative western blot of p-Smad2, p-Smad3 and t-Smad2/3. (F, G) Western blot analysis of p-Smad2/t-Smad2 and p-Smad3/t-Smad3. Values are the mean ± SD, and western blots were conducted at least 3 times. *P < 0.05; ***P < 0.001 vs. control; ††P < 0.01 vs. vehicle.

Article Snippet: The blocking was done with 5% nonfat milk for 2 h, and then membranes were incubated with different rabbit anti-rat primary antibodies, including ALK7 antibody (R&D, Minneapolis, MN, USA), phosphor-Akt (Ser473)/Akt, phosphor-Smad2 (Ser465/467)/Smad2, phosphor-Smad3 (Ser423/425)/Smad3, Bcl-2, Bax and cleaved Caspase3 antibody (Cell Signaling Technology, Beverly, MA, USA), and Collagen I and III antibody (Abcam, Cambridge, UK) overnight at 4°C.

Techniques: Phospho-proteomics, Expressing, Western Blot, Control