Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Myostatin propeptide mutation of the hypermuscular Compact mice decreases the formation of myostatin and improves insulin sensitivity.

doi: 10.1152/ajpendo.00216.2016

Figure Lengend Snippet: Fig. 3. Myostatin level in skeletal muscle of Compact, congenic wild-type, and BALB/c mice. M. gastrocnemius protein extracts were subjected to SDS-PAGE and blotted with anti-myostatin or anti-propeptide antibody. Represen- tative images are shown. Note the presence of mature myostatin dimer and myostatin propeptide in Compact sam- ples. Mouse recombinant myostatin was used as a positive control, and muscle homogenates of myostatin knockout (KO) mice served as a negative control. Differences in glycosylation may cause altered electrophoretic mobility. Bar diagrams show the quantification of the results. Data are reported as means SE; n 5 Compact, 5 congenic wild-type, and 6 BALB/c mice. *P 0.05; **P 0.01; ***P 0.001.

Article Snippet: Mouse recombinant myostatin (788-G8; R&D Systems) was used as a positive control.

Techniques: SDS Page, Recombinant, Positive Control, Knock-Out, Negative Control, Glycoproteomics

Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Myostatin propeptide mutation of the hypermuscular Compact mice decreases the formation of myostatin and improves insulin sensitivity.

doi: 10.1152/ajpendo.00216.2016

Figure Lengend Snippet: Fig. 5. Glucose tolerance and insulin sensitivity are improved by Compact myostatin mutation and reduced in congenic wild-type mice. Intraperitoneal (ip) glucose tolerance (A and B) and insulin sensitivity tests (C and D) of 3- to 4-mo-old (A and C) and 10-mo-old animals (B and D). Area under the curve (AUC) values are presented in bar diagrams. Data are reported as means SE. *P 0.05 and **P 0.01; n 3 Compact, 7 congenic wild-type, and 3 BALB/c mice (A), n 7 Compact, 3 congenic wild-type, and 6 BALB/c mice (B), n 3 Compact, 3 congenic wild-type, and 4 BALB/c mice (C), and n 6 Compact, 4 congenic wild-type, and 4 BALB/c mice (D).

Article Snippet: Mouse recombinant myostatin (788-G8; R&D Systems) was used as a positive control.

Techniques: Mutagenesis

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Angiotensin (1-7) (Ang-(1-7)) prevented the myostatin-induced decrease of the myotube diameter. ( A ) C 2 C 12 myoblasts differentiated for five days (myotubes) were pre-incubated in the absence or presence of Ang-(1-7) (10 nM) for 60 min, and then incubated with myostatin (Mstn, 1 µg/mL) for 72 h. Myosin heavy chain (MHC; green) was detected through indirect immunofluorescence (IFI). Hoechst was used to stain the nuclei (blue). The bar scale represents 100 μm. ( B ) The graphics show the distribution of the myotube diameters. The values are expressed as a percentage of the total myotubes and correspond to the mean ± standard deviation (SD) from three independent experiments ( n = 3; * p < 0.05 vs. control).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Incubation, Immunofluorescence, Staining, Standard Deviation, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Ang-(1-7) decreased the myostatin-dependent decline in the myofibrillar protein levels in the myotubes. The C 2 C 12 myotubes were incubated with myostatin (Mstn, 1 µg/mL) for 72 h in the presence or absence of Ang-(1-7) (10 nM; pre-incubated for 1 h). ( A ) The MHC, troponin, and β-actin protein levels were determined by Western blot analysis. The molecular weights are shown in kDa. Densitometric analysis for the MHC ( B ) and troponin ( C ) protein levels with values normalized to β-actin and expressed as a fold of change relative to the control (myotubes without treatment). All of the values correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Incubation, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Ang-(1-7) avoided the myostatin-dependent induction of atrogin-1 and MuRF-1 in the myotubes. The C 2 C 12 cells differentiated for five days were pretreated or not with Ang-(1-7) (10 nM) for 1 h, and further with myostatin (Mstn, 1 µg/mL) for 12 or 24 h. The mRNA levels of atrogin-1 ( A ) and MuRF-1 ( B ) were determined by RT-qPCR. ( C ) The protein levels of atrogin-1 and MuRF-1 were detected by Western Blot analysis. β-actin was used as the loading control. The molecular weights are shown in kDa. Quantitative analysis of atrogin-1 ( D ) and MuRF-1 ( E ) levels with values normalized to β-actin. All of the values correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Quantitative RT-PCR, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Ang-(1-7) reduced the reactive oxygen species (ROS) production induced by myostatin in the myotubes. ( A ) C 2 C 12 myotubes were incubated with myostatin (Mstn, 1 µg/mL) for 24 h in the presence or absence of Ang-(1-7) (10 nM; pre-incubated for 1 h). The ROS levels were then determined by the detection of fluorescence using a CM-DCF-DA probe. The nuclei were labelled with Hoechst. The scale bar represents 100 µm. ( B ) Quantification of the DCF-positive myotubes. The values are expressed as a fold of induction relative to the myotubes without treatment, and correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Incubation, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Ang-(1-7) reduced the myostatin-induced NF-κB signaling in myotubes. ( A ) C 2 C 12 myotubes were pre-incubated with or without Ang-(1-7) (10 nM) and after 1 h with myostatin (Mstn, 1 µg/mL) for 24 h. Protein levels of IκB and β-actin were detected by Western blot analysis. Molecular weights are shown in kDa. ( B ) Quantitative analysis for IκB protein levels with values normalized to β-actin. ( C ) Luciferase activity of pNF-κB-luc. ( D ) The mRNA levels of TNF-α were determined by RT-qPCR. The values are expressed as a fold of induction relative to the control (myotubes without treatment), and correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Incubation, Western Blot, Luciferase, Activity Assay, Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Reduction of myostatin dependent on muscle atrophy and signaling by Ang-(1-7) is mediated through Mas receptor in myotubes. C 2 C 12 myotubes, pretreated with or without Ang-(1-7) (10 nM), A779 (10 µM) (pre-incubated for 30 min), or both, were incubated with myostatin (Mstn, 1 µg/mL) for 12 or 24 h. The parameters measured were: ( A ) distribution of the myotube diameters; mRNA levels of ( B ) atrogin-1, ( C ) MuRF-1, and ( E ) TNF-α; ( D ) Luciferase activity of pNF-κB-luc; and ( F ) ROS levels. The values are expressed as a fold of induction relative to the myotubes without treatment and correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Incubation, Luciferase, Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin (1-7) Decreases Myostatin-Induced NF-κB Signaling and Skeletal Muscle Atrophy

doi: 10.3390/ijms21031167

Figure Lengend Snippet: Prevention of myostatin-dependent muscle atrophy and signaling by Ang-(1-7) is mediated by the Akt/PKB activity in the myotubes. The C 2 C 12 myotubes were incubated with myostatin (Mstn, 1 µg/mL) for 12 or 24 h in the presence or absence of Ang-(1-7) (10 nM), MK2206 (10 µM; pre-incubated for 30 min), or both. The parameters measured were as follows: ( A ) distribution of the myotube diameters; mRNA levels of ( B ) atrogin-1, ( C ) MuRF-1 and ( E ) TNF-α; ( D ) Luciferase activity of pNF-κB-luc; and ( F ) ROS levels. The values are expressed as a fold of induction relative to the myotubes without treatment, and correspond to the mean ± SD from three independent experiments ( n = 3; * p < 0.05 vs. control without treatment).

Article Snippet: The myotubes were incubated with 1 μg/ml of human recombinant myostatin (Cell Sciences Inc., MA, USA) and/or 10 nM of Ang-(1-7) (Sigma-Aldrich, USA), as indicated in each figure.

Techniques: Activity Assay, Incubation, Luciferase, Control