Journal: Scientific Reports

Article Title: Loss of microRNA-23–27–24 clusters in skeletal muscle is not influential in skeletal muscle development and exercise-induced muscle adaptation

doi: 10.1038/s41598-018-37765-3

Figure Lengend Snippet: Skeletal muscle phenotype of miR-23–27–24-cluster-dKO mice. ( A ) Representative images of western blotting analysis of plantaris muscle. ( B ) Quantitative data for the protein expression of MyHC type IIa, MyHC type IIb, PGC-1α, and COX IV. Data are means ± SEM (n = 6). ( C ) Representative images of western blotting analysis of soleus muscle. ( D ) Quantitative data for the protein expression of MyHC type I, MyHC type IIa, PGC-1α, and COX IV. Data are means ± SEM (n = 5–6). ( E ) Representative images of plantaris and soleus muscles immunofluorescently stained for MyHC type IIa fibers and dystrophin in green, and type IIb fibers in red. MyHC type IId/x fibers are unstained (black). Scale bar, 50 μm. ( F ) Proportions of muscle fiber types in plantaris and soleus muscles. Proportion of each fiber type is presented as a percentage. Muscle fibers in entire cross-sections were counted. I, MyHC type I; IIa, MyHC type IIa; IIb, MyHC type IIb; IId/x, MyHC type IId/x. Data are means ± SEM (n = 4–5). # P < 0.05 compared with WT. ( G ) Total fiber numbers in plantaris and soleus muscles. Muscle fibers in entire cross-sections were counted. Data are means ± SEM (n = 4–5). ( H ) Distributions of MyHC type IIa fibers (top) MyHC type IIb fibers (middle), and MyHC type IId/x fibers (bottom) in plantaris muscle. Graphs show percentages of fibers per cross-sectional area (CSA). Data are means ± SEM (n = 5). ( I ) CSA distributions of MyHC type I fibers (top), MyHC type IIa (middle), and MyHC type IId/x fibers (bottom) in soleus muscle. Data are means ± SEM (n = 5). ( J ) mRNA expression of Myostatin, MurF1 and Atrogin-1 in WT and dKO mice skeletal muscle. Data are means ± SEM (n = 6). K, Quantitative data for the phospho-Akt/Akt by western blotting in WT and dKO mice skeletal muscle. Data are means ± SEM (n = 6).

Article Snippet: The membranes were incubated with the following primary antibodies: anti-myosin heavy chain (MyHC) type I (BA-F8, DSMZ, 1:100), anti-MyHC type IIa (SC-71, DSMZ, 1:100), anti-MyHC type IIb (BF-F3, DSMZ, 1:100), anti-PGC1-α (AB3242, Millipore, 1:500), anti-cytochrome c oxidase complex IV (anti-COX IV; #4844, Cell Signaling Technology, 1:1000), anti-CD31 (anti-PECAM-1; sc-46694, Santa Cruz Biotechnology, INC., 1:400), anti-Phospho-Akt (#4060, Cell signaling Technology, 1:1000), anti-Akt (#9272, Cell signaling Technology, 1:1000) and anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH; MAB374, Millipore, 1:1000).

Techniques: Western Blot, Expressing, Muscles, Staining

Journal: Scientific Reports

Article Title: Loss of microRNA-23–27–24 clusters in skeletal muscle is not influential in skeletal muscle development and exercise-induced muscle adaptation

doi: 10.1038/s41598-018-37765-3

Figure Lengend Snippet: miR-23–27–24 clusters are not essential for endurance-exercise-induced muscle adaptation. ( A ) Representative images of western blotting analysis. ( B ) Quantitative data for the protein expression of MyHC type IIa, MyHC type IIb, PGC-1α, and COX IV. Data are means ± SEM (n = 6). **P < 0.01 main effect of exercise. ( C ) Representative images of SDH staining in plantaris muscle. Scale bar, 100 μm. ( D ) Representative immunofluorescent images of capillaries in plantaris muscle. Dystrophin is shown in red, CD31 in green. Scale bar, 100 μm. Quantitative data for the capillary-to-fiber ratio ( E ) and CD31 expression ( F ) in plantaris muscle. Data are means ± SEM (n = 5). **P < 0.01 main effect of exercise. ( G ) Representative images of plantaris muscle immunofluorescently stained for MyHC type IIa fibers and dystrophin in green, type IIb fibers in red. MyHC type IId/x fibers are unstained (black). Scale bar, 500 μm. ( H ) Mean cross-sectional areas (CSAs) of MyHC type IIa, MyHC type Id/x, and MyHC type IIb fibers in plantaris muscle. Data are means ± SEM (n = 4–5). # P < 0.05 main effect of genotype. **P < 0.01 main effect of exercise. ( I ) Proportions of MyHC IIa, MyHC IId/x, and MyHC IIb fibers in plantaris muscle. Data are means ± SEM (n = 4). # P < 0.05 main effect of genotype. **P < 0.01 main effect of exercise. ( J ) Expressions of the mature miRNAs in miR-23a/b clusters after 4 weeks of voluntary running. Data are means ± SEM (n = 5). ## P < 0.01 main effect of genotype. *P < 0.05 main effect of exercise. sed, sedentary group; ex, exercised group.

Article Snippet: The membranes were incubated with the following primary antibodies: anti-myosin heavy chain (MyHC) type I (BA-F8, DSMZ, 1:100), anti-MyHC type IIa (SC-71, DSMZ, 1:100), anti-MyHC type IIb (BF-F3, DSMZ, 1:100), anti-PGC1-α (AB3242, Millipore, 1:500), anti-cytochrome c oxidase complex IV (anti-COX IV; #4844, Cell Signaling Technology, 1:1000), anti-CD31 (anti-PECAM-1; sc-46694, Santa Cruz Biotechnology, INC., 1:400), anti-Phospho-Akt (#4060, Cell signaling Technology, 1:1000), anti-Akt (#9272, Cell signaling Technology, 1:1000) and anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH; MAB374, Millipore, 1:1000).

Techniques: Western Blot, Expressing, Staining

Journal: PLoS ONE

Article Title: Decrease in Blood Pressure and Regression of Cardiovascular Complications by Angiotensin II Vaccine in Mice

doi: 10.1371/journal.pone.0060493

Figure Lengend Snippet: (A) The mouse heart weights are shown. The results are expressed as the mean heart weight per body weight ± the standard error of the mean. (B) The cardiac tissue was stained with Masson trichrome and analyzed for cardiac fibrosis. Representative photographs are shown. The scale bar represents 20 µm. The results are expressed as the mean of the fibrotic area ± SE of the mean. (C) The mRNA levels of collagen type I (collagen I), collagen type III (collagen III), atrial natriuretic factor (ANF) and β-myosin heavy chain (β-MHC) were evaluated using RT-PCR. The results are expressed as the ratio of gene expression in the experimental mice to gene expression in the control mice ± SE of the mean. Control mice were immunized with KLH (1 mg, with adjuvant) and experimental mice were immunized with Ang II-KLH (1,000 ng, with adjuvant). All groups contained 6 to 8 mice on day 56. * P <0.05.

Article Snippet: The relative gene copy numbers were quantified with real-time PCR using TaqMan Gene Expression Assays (18S: 4352930; collagen type I: Mm00801606; collagen type III: Mm01254476; ANF: Mm01255748; and β-MHC: Mm00600555; Applied Biosystems, Foster City, CA).

Techniques: Staining, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Control, Adjuvant