Journal: Biochemistry and Biophysics Reports

Article Title: Beta-agonist drugs modulate the proliferation and differentiation of skeletal muscle cells in vitro

doi: 10.1016/j.bbrep.2021.101019

Figure Lengend Snippet: Effect of β-agonists on myogenic proteins expression during cell differentiation. (A) Western blot imaging of proteins extracted at differentiation days 0, 1, 4, and 7 from cells exposed to drugs at seeding. Densitometry analysis of (B) MyHC, (C) α-actinin-2, (D) MyoD, and (E) myogenin proteins normalized against GAPDH. Data are presented as mean ± SEM of three independent biological replicates. Ct: control, Cl: clenbuterol, Sa: salbutamol, Ci: cimaterol, Ra: ractopamine. (*) p < 0.05.

Article Snippet: The membranes were then blocked with 5% non-fat milk for 1 h and first antibodies of Heavy Chain Cardiac Myosin (Genetex, GTX20015, 1: 1000), alpha Actinin 2 (Genetex, GTX103219, 1:3000), Myogenin (5FD) (Novus, NB100-56510, 1:1500), MyoD1 (phosphor Ser200) (Genetex, GTX50144, 1:500), and GAPDH (Genetex, GTX100118, 1: 100 000) were applied for overnight at 4 °C.

Techniques: Expressing, Cell Differentiation, Western Blot, Imaging, Control

Journal: Neural Regeneration Research

Article Title: Transferrin receptor 1 plays an important role in muscle development and denervation-induced muscular atrophy

doi: 10.4103/1673-5374.301024

Figure Lengend Snippet: Severe muscular atrophy in HSA-Cre;Tfr1 F/F mutant mice. (A) Hematoxylin-eosin staining of the cross-sections of gastrocnemius muscle in HSA-Cre; Tfr1 F/F mutants and control littermates at P10. The outline of a single muscle fiber is indicated by a dashed line, revealing smaller cross-sectional area of muscle fibers in mutants compared with controls. Scale bars: 30 μm. (B, C) Quantitative analysis of the average muscle fiber area shows a significant reduction in HSA-Cre;Tfr1 F/F mutants compared with control littermates at P10, but the number of muscle fibers was nearly the same. (D) Ultrastructural analysis of gastrocnemius muscles by electron microscopy. No significant misalignment of Z-lines (arrows) or central nuclei was found in either HSA-Cre;Tfr1 F/F mutants or control littermates. Scale bars: 500 nm (left), 100 nm (right). (E) Immunofluorescence staining of gastrocnemius muscle cross-sections labeled with anti-myosin (slow) (arrowheads) or anti-myosin (fast) (asterisks) antibody (red, Alexa Fluor 568) in HSA-Cre;Tfr1 F/F mutants and control littermates. Increased expression of slow-type myosin, but reduced expression of fast-type myosin, was found in HSA-Cre;Tfr1 F/F mutants compared with control littermates. Scale bar: 100 mm. (F) Representative western blot shows the expression of TfR1, myosin (slow), myosin (fast), MyoD and myogenin in skeletal muscles from HSA-Cre;Tfr1 F/F mutants and control littermates at P10. (G) Quantitative analysis of western blots showing significantly decreased expression of TfR1, myosin (fast), MyoD and myogenin, but increased expression of myosin (slow), in HSA-Cre;Tfr1 F/F mutants compared with control littermates. (H) Quantitative reverse transcription-polymerase chain reaction analysis indicating significantly downregulated expression of Myod1 , Myogenin , Mef2c and MCK genes in HSA-Cre;Tfr1 F/F mutant mice. In contrast, Foxo1 was significantly upregulated in HSA-Cre;Tfr1 F/F mutants compared with control littermates. Data are expressed as the mean ± SEM ( n = 6). * P < 0.05, ** P < 0.01 (Student’s t -test). Foxo1: Forkhead box protein O1; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; HSA: skeletal muscle actin; MCK: muscle creatine kinase; Mef2C: myocyte enhancer factor 2C; n.s.: not significant; P10: postnatal day 10; Tfr1: transferrin receptor 1.

Article Snippet: The following antibodies were used: rabbit anti-TfR1 antibody (1:500; Cat# 226290; Biocompare, Salem, MA, USA), mouse anti-myosin (skeletal, fast) antibody (1:1000; Cat# M4276; Sigma-Aldrich), mouse anti-myosin (skeletal, slow) antibody (1:1000; Cat# M8421; Sigma-Aldrich), rabbit anti-MyoD antibody (a muscle transcriptional regulatory factor; 1:1000; Cat# MBS856897; Biocompare, San Diego, CA, USA), goat anti-MyoG antibody (a muscle transcriptional regulatory factor; 1:1000; Cat# SAB2501587; Sigma-Aldrich), mouse anti-glyceraldehyde 3-phosphate dehydrogenase antibody (1:2000; Cat# KM9002T; Sungenebiotech, Tianjin, China).

Techniques: Mutagenesis, Staining, Electron Microscopy, Immunofluorescence, Labeling, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Science Progress

Article Title: Preventive effect and possible mechanisms of ultrashort wave diathermy on myogenic contracture in a rabbit model

doi: 10.1177/00368504211054992

Figure Lengend Snippet: Protein levels of myogenic differentiation (MyoD) and muscle atrophy F-box (MAFbx-1) relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). (A) Photographs of MyoD, MAFbx-1 and GAPDH protein-band intensities. (B) MyoD protein levels relative to GAPDH. (C) MAFbx-1 protein levels relative to GAPDH. a p < 0.05 vs. Group C(I-0), b p < 0.05 vs. Group I-1, c p < 0.05 vs. Group I-2, a’ p < 0.05 vs. Group I-0, b’ p < 0.05 vs. Group U-1, c ’ p < 0.05 vs. Group U-2, *** p < 0.001, ** p < 0.01 and * p < 0.05.

Article Snippet: Subsequently, the membranes were incubated with rabbit anti-MyoD antibody (1:5000; Beyotime), rabbit anti-MAFbx-1 antibody (1:2000; Beyotime) and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (1:8000; Beyotime) at 4 °C for 12 h. The following day, the membranes were washed in TBST five times for 3 min and incubated with peroxidase-conjugated affinity-purification goat anti-rabbit IgG-HRP (1: 10,000; Beyotime), as the secondary antibody, for 1 h at 20–25 °C.

Techniques: