Journal: Sports Medicine and Health Science

Article Title: Exercise preconditioning prevents immobilization-induced skeletal muscle atrophy by activating Prmt1-p38/ATF2-Sesn1 signaling axis in C57BL/6J mice

doi: 10.1016/j.smhs.2025.04.001

Figure Lengend Snippet: Exercise preconditioning prevented muscle atrophy through protein arginine methyltransferase 1 (Prmt1)-Sestrin1 (Sesn1)-transcriptional co-activator PPAR-γ co-activator-1 α (PGC-1α)-mediated skeletal muscle regeneration. (A–B) Western blot results of MyoD, Myf5, MEF2 and MyoG in GAS muscle. (C–D) Co-IP results, IP: MyoD, GAS muscle was used. A-B, ∗ p ​< ​0.05 vs. C; ∗∗ p ​< ​0.01 vs. C; ## p ​< ​0.01 vs. Im; $ p ​< ​0.05 vs. E ​+ ​Im; $$ p ​< ​0.01 vs. E ​+ ​Im. Two-way ANOVA was used, and data are shown as means ​± ​standard error of the mean ( SEM ) (C, n ​= ​8, Im, n ​= ​8, E ​+ ​Im, n ​= ​8, E+5003+Im, n ​= ​8). (E–F) H&E staining of myotubes at each differentiation time points of C2C12 myoblasts overexpressing Sesn1. Scale bar ​= ​100 ​μm ∗ p ​< ​0.05 vs. Ad-Sesn1-, unpaired Student's t-test was used and data are shown as means ​± ​ SEM ( n ​= ​3 in each group). (G–H) Western blot results of Sesn1, Prmt1, PGC-1α, MyoD, MEF2, Myf5, and MyoG in C2C12 myoblasts at each time points of differentiation. ∗ p ​< ​0.05 vs. D0, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (I–J) Western blot results of PGC-1α in nucleus of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group). (K–L) Western blot results of PGC-1α in cytoplasm of C2C12 myoblasts overexpressing Sesn1 at each time points of differentiation. ∗∗ p ​< ​0.01 vs. Ad-Sesn1-, unpaired Student's t -test was used and data are shown as means ​± ​ SEM ( n ​= ​6 in each group).

Article Snippet: The antibodies are listed below: aDMA (Anti-Asymmetric Di-Methyl Arginine Motif) (1:1 500, Rabbit, Cell Signal Tech, USA), Akt (protein kinase B) (1:2 000, Mouse, Proteintech, USA), pAkt-Ser473 (1:2 000, Rabbit, Cell Signal Tech, USA), AMPKα2 (1:2 000, Rabbit, Cell Signal Tech, USA), pAMPK-Thr172 (1:2 000, Rabbit, Cell Signal Tech, USA), ATF2 (1:2 000, Rabbit, Proteintech, USA), Atrogin-1 (FBXO32) (1:15 000, Mouse, Proteintech, USA), FoxO3a (Forkhead box O3) (1:1 000, Mouse, Proteintech, USA), pFoxO3a-Ser315 (1:2 000, Rabbit, Proteintech, USA), GAPDH (1:5 000, Rabbit, Utibody, CN), IGF-1 (insulin-like growth factor 1) (1:2 000, Mouse, Proteintech, USA), LaminB (1:2 000, Rabbit, Abcam, USA), MEF2 (myocyte enhancer factor 2) (1:2 000, Rabbit, Proteintech, USA), Myf5 (myogenic factor 5) (1:2 000, Rabbit, Abclonal, CN), MyoD (myogenic differentiation antigen) (1:2 000, Rabbit, Proteintech, USA), MyoG (myogenin) (1:2 000, Rabbit, Proteintech, USA). mTOR (mammalian target of rapamycin) (1:2 000, Rabbit, Cell Signal Tech, USA), p38 (1:2 000, Rabbit, WANLEIBIO, CN), p-p38-Thr180/Tyr182 (1:2 000, Rabbit, WANLEIBIO, CN), PGC-1α (1:1 000, Rabbit, Abcam, USA), Prmt1 (1:2 000, Rabbit, Cell Signal Tech, USA), Raptor (1:1 000, Rabbit, Cell Signal Tech, USA), Sesn1 (1:1 000, Rabbit, HUABIO, CN), Sesn1 (1:1 000, Rabbit, Abcam, USA), TRIM63 (MuRF1) (1:2 000, Rabbit, Proteintech, USA).

Techniques: Western Blot, Co-Immunoprecipitation Assay, Staining

Journal: Nutrients

Article Title: Ophiopogon japonicus Root Extract Attenuates Obesity-Induced Muscle Atrophy Through Regulation of the PI3K-AKT-mTOR/FoxO3a Signaling Pathway and Lipid Metabolism in Mice and C2C12 Myotubes

doi: 10.3390/nu17243946

Figure Lengend Snippet: Effects of OJ extract on PA-induced atrophy in C2C12 myotubes. ( A ) Morphological change in C2C12 myotubes after treatment of PA at different concentrations for 48 h. ( B ) Protein expression of MyHC, Myogenin, Atrogin-1, and MuRF1 in C2C12 myotubes with PA treatment. ( C ) Representative images of MyHC and Myogenin immunofluorescence in C2C12 myotubes (Scale bar = 100 μm). ( D ) Expression of MyHC, Myogenin, Atrogin-1, and MuRF1 in C2C12 myotubes after treatment of OJ extract at different concentrations with PA at 0.5 mM. All data are presented as the mean ± SD ( n = 3). The p values were defined as follows: # p < 0.05, ## p < 0.01 and ### p < 0.001 vs. CON; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. PA.

Article Snippet: The membranes were incubated in 5% non-fat dry milk for 2 h at RT to block nonspecific binding and subsequently incubated overnight at 4 °C with the respective primary antibodies, including MyHC (sc-376157, Santa Cruz), Myogenin (sc-52903), MuRF1 (bs-2539R, Bioss Antibodies, Woburn, MA, USA), Atrogin-1 (PA5-91959, Thermo Fisher Scientific), p -FoxO3a (PA5-36816), FoxO3a (PA5-27145), SREBP-1c (PA1-337), CPT1b (PA5-79065), mTOR (2972s, Cell Signaling, Danvers, MA, USA), p -mTOR (5536s), AKT (9272s), p -AKT (9271s), PI3K (4292s), p -PI3K (4228s), β-Actin (A5316, Sigma-Aldrich), and GAPDH (PA0018).

Techniques: Expressing, Immunofluorescence

Journal: The American Journal of Pathology

Article Title: Strong Immunostaining for Myogenin in Rhabdomyosarcoma Is Significantly Associated with Tumors of the Alveolar Subclass

doi: 10.1016/s0002-9440(10)64743-8

Figure Lengend Snippet: Figure 1. Western blot analysis of myogenin expression in various small round cell tumor lines using the anti-myogenin monoclonal antibody. The antibody only reacts with a protein band corresponding to the molecular mass of myogenin (approximately 34 kd) in the Rh30 rhabdomyosarcoma cell lysate. All other small round cell tumor lysates were negative. Rh30, alveolar rhabdomyosarcoma cell line; PFSK-1A, primitive neuroectodermal tumor cell line; EB2, lymphoma cell line; SKNSH, neuroblastoma cell line; SJSA-1, Ewing’s sarcoma cell line.

Article Snippet: The monoclonal antibody to myogenin (Imgenex, San Diego, CA) was raised against glutathione S-transferase (GST)-myogenin fusion protein.

Techniques: Western Blot, Expressing

Journal: The American Journal of Pathology

Article Title: Strong Immunostaining for Myogenin in Rhabdomyosarcoma Is Significantly Associated with Tumors of the Alveolar Subclass

doi: 10.1016/s0002-9440(10)64743-8

Figure Lengend Snippet: Figure 2. Representative staining for myogenin in embryonal and alveolar rhabdomyosarcoma. The embryonal rhabdomyosarcoma (top panels) in this figure was less than 10% positive for myogenin and was scored as (), whereas most tumor cells (85–95%) in the alveolar rhabdomyosarcoma tumor (bottom panels) were homogeneously strongly positive for myogenin. This tumor was scored as (). Left: Lower magnification (original magnification, 200). Right: Higher magnification (original magnification, 400). ABC staining, methyl green counterstain.

Article Snippet: The monoclonal antibody to myogenin (Imgenex, San Diego, CA) was raised against glutathione S-transferase (GST)-myogenin fusion protein.

Techniques: Staining

Journal: The American Journal of Pathology

Article Title: Strong Immunostaining for Myogenin in Rhabdomyosarcoma Is Significantly Associated with Tumors of the Alveolar Subclass

doi: 10.1016/s0002-9440(10)64743-8

Figure Lengend Snippet: Figure 3. Western blot analysis for myogenin expression in a smaller subset of rhabdomyosarcoma specimens. Four of six alveolar rhabdomyosarcomas (lanes 8, 10, 11, and 12) were strongly positive for myogenin. One alveolar rhabdomyosarcoma (lane 7) was negative, and one alveolar tumor (lane 9) was weakly positive for myogenin. Three of six embryonal rhabdomyosar- comas (lanes 3, 4, and 6) were negative for myogenin, whereas two of six (lanes 2 and 5) were weakly positive. One embryonal rhabdomyosarcoma (lane 1), a posttreatment tumor, was strongly positive for myogenin. The blot was incubated with anti-myogenin monoclonal antibody. Reactivity was detected using a horseradish peroxidase-conjugated secondary antibody and visualized using a chemiluminescent substrate and exposure to photographic films.

Article Snippet: The monoclonal antibody to myogenin (Imgenex, San Diego, CA) was raised against glutathione S-transferase (GST)-myogenin fusion protein.

Techniques: Western Blot, Expressing, Incubation

Journal: eLife

Article Title: A novel mouse model for LAMA2 -related muscular dystrophy with analysis of molecular pathogenesis and clinical phenotype

doi: 10.7554/eLife.94288

Figure Lengend Snippet: ( A ) Immunofluorescence staining for myosin heavy chain (MYHC), desmin, β-tubulin, MYOG, MYOD1, and MYF-5 in WT and dy H /dy H muscles at P14. MYHC, MYOG, MYOD1, and MYF-5 were focally increased, while desmin and β-tubulin showed mild decrease in the cytoplasm of hypertrophic muscle fibers as well as focal increase in developmental muscle fibers in dy H /dy H mice. Wheat-germ agglutinin (WGA; red) was used to visualize the muscle fibers and connective tissue. White asterisk indicated focal regions with changes. ( B ) F-actin, α-actin, MYH2, MYHC, desmin, β-tubulin, MYOG, and MYOD1 were detected by Western blot in P14 WT (n=6) and dy H /dy H (n=6) muscles. The levels of F-actin, MYOG and MYOD1 were significantly decreased in dy H /dy H muscles (p<0.05), MYH2 and MYHC were significantly increased in dy H /dy H muscles (p<0.05), while α-actin, desmin, and β-tubulin showed no significant difference between WT (n=6) and dy H /dy H (n=6) mice. Figure 8—source data 1. Original files for western blot analysis displayed in . Figure 8—source data 2. PDF file containing original western blots for , indicating the relevant bands.

Article Snippet: The antibodies used in this study were as follows: laminin α2 chain (rat monoclonal, 4H8-2, Sigma, Saint Louis, USA), MYOD1 (mouse monoclonal, ab64159, Abcam, Cambridge, UK), MYOG (mouse monoclonal, MAB66861, R&D Systems, Minneapolis, USA), myosin heavy chain 2 (MYH2) (rabbit monoclonal, ab124937, Abcam, Cambridge, UK), MYHC (mouse monoclonal, MAB4470, R&D Systems, Minneapolis, USA), desmin (mouse monoclonal, MA5-15306, Invitrogen, CA), α-actin (rabbit polyclonal, 23660-I-AP, Proteintech, Rosemont, USA), F-actin (rabbit polyclonal, bs-1571R, Bioss, Rosemont, USA).

Techniques: Immunofluorescence, Staining, Muscles, Western Blot

Journal: eLife

Article Title: A novel mouse model for LAMA2 -related muscular dystrophy with analysis of molecular pathogenesis and clinical phenotype

doi: 10.7554/eLife.94288

Figure Lengend Snippet: ( A ) Quantitative analysis of immunofluorescence staining for myosin heavy chain (MYHC), MYOG, desmin, MYOD1, β-tubulin, and MYF-5 in wild-type (WT) and dy H /dy H muscles at P14. ( B ) Quantitative analysis of Western blot for laminin α2, F-actin, α-actin, MYH2, MYHC, desmin, β-tubulin, MYOG, and MYOD1 in P14 WT and dy H /dy H muscles. *** for p-value <0.001, ** for p-value <0.01, * for p-value <0.05.

Article Snippet: The antibodies used in this study were as follows: laminin α2 chain (rat monoclonal, 4H8-2, Sigma, Saint Louis, USA), MYOD1 (mouse monoclonal, ab64159, Abcam, Cambridge, UK), MYOG (mouse monoclonal, MAB66861, R&D Systems, Minneapolis, USA), myosin heavy chain 2 (MYH2) (rabbit monoclonal, ab124937, Abcam, Cambridge, UK), MYHC (mouse monoclonal, MAB4470, R&D Systems, Minneapolis, USA), desmin (mouse monoclonal, MA5-15306, Invitrogen, CA), α-actin (rabbit polyclonal, 23660-I-AP, Proteintech, Rosemont, USA), F-actin (rabbit polyclonal, bs-1571R, Bioss, Rosemont, USA).

Techniques: Immunofluorescence, Staining, Muscles, Western Blot

Journal: Life Science Alliance

Article Title: SMN promotes mitochondrial metabolic maturation during myogenesis by regulating the MYOD-miRNA axis

doi: 10.26508/lsa.202201457

Figure Lengend Snippet: Resources and reagents used in this study.

Article Snippet: Myogenin antibody (MGN185) Alexa Fluor 647 , NOVUS , Cat# NBP2-33056AF647.

Techniques: Transduction, Control, Negative Control, Virus, Expressing, Construct, Recombinant, Flow Cytometry, Isolation, Software

Journal: PLoS ONE

Article Title: Network Analysis for the Identification of Differentially Expressed Hub Genes Using Myogenin Knock-down Muscle Satellite Cells

doi: 10.1371/journal.pone.0133597

Figure Lengend Snippet: A) The hub CTNNA2 (yellow) and its first neighbors. Genes that were up-regulated in MYOG kd cells are shown in green whereas the down-regulated genes are shown in red. The genes predicted by GeneMania are shown in cyan. B) Functional enrichment of first neighbors of CTNNA2. The green circles represent up-regulated gene nodes and are enriched in metabolic processes. The larger red circles represent down-regulated nodes and are enriched in processes related to cell growth, morphogenesis, and migration. The smallest circle (cyan) represents the group of genes predicted by GeneMania and is enriched for DNA replication and DNA metabolic processes.

Article Snippet: C2C12 cells grown to 30% confluence were transfected with 1 ng of vector containing MYOG shRNA or the scrambled vector using transfection reagent (Santa Cruz Biotechnology, CA, USA).

Techniques: Functional Assay, Migration

Journal: PLoS ONE

Article Title: Network Analysis for the Identification of Differentially Expressed Hub Genes Using Myogenin Knock-down Muscle Satellite Cells

doi: 10.1371/journal.pone.0133597

Figure Lengend Snippet: Bovine MSCs were cultured for 10, 12, 14, and 16 days in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin (P/S). RNA was then isolated. A) CTNNA2 expression was analyzed by real-time RT-PCR and had increased on Day 12 compared to Day 10. B) MYOG gene expression was knocked down in bovine MSCs. MYOG and CTNNA2 expression was measured in MYOG knock-down cells. C2C12 cells were cultured with differentiation media for 0, 2, 4, and 6 days. mRNA expression was then analyzed by real-time RT-PCR. C) CTNNA2 expression was elevated on Day 6 compared to Day 0. D) Protein localization was observed by immunocytochemistry on Day 0 and 6. CTNNA2 protein was highly expressed inside myotubes on Day 6. E) MYOG expression was knocked down with shRNA in C2C12 cells that were cultured in differentiation media for 6 days. Gene expression was analyzed by real-time RT-PCR. Expression of MYOG and CTNNA2 was down-regulated in MYOG kd . The p-value indicated statistical significance of the data (p<0.05, mean ± standard deviation [S.D.], n = 3).

Article Snippet: C2C12 cells grown to 30% confluence were transfected with 1 ng of vector containing MYOG shRNA or the scrambled vector using transfection reagent (Santa Cruz Biotechnology, CA, USA).

Techniques: Cell Culture, Isolation, Expressing, Quantitative RT-PCR, Gene Expression, Knockdown, Immunocytochemistry, shRNA, Standard Deviation

Journal: PLoS ONE

Article Title: Network Analysis for the Identification of Differentially Expressed Hub Genes Using Myogenin Knock-down Muscle Satellite Cells

doi: 10.1371/journal.pone.0133597

Figure Lengend Snippet: C2C12 cells were transfected with CTNNA2-specific siRNA and incubated with 2% FBS for 6 d. A) Morphology of the CTNNA2 wt and CTNNA2 kd cells was similar. B) The fusion index was determined and the values were similar for CTNNA2 wt and CTNNA2 kd cells. C) mRNA expression was analyzed by real-time RT-PCR. CTNNA2 expression was decreased in CTNNA2 knock-down cells. However, MYOG expression was similar in the CTNNA2 wt and CTNNA2 kd cells . D) Expression of ECM genes was analyzed by real-time RT-PCR. COL1α1, COL1α2, and MSTN gene expression was increased in CTNNA2 kd cells. In particular, MSTN expression was the highest among the four genes. E) Protein expression of CTNNA2, COL1α1, and MSTN in CTNNA2 kd was analyzed by Western blotting. CTNNA2 and ECM protein expression levels were similar to the mRNA expression patterns. The p-value indicates the statistical significance (p<0.05, mean ± S.D., n = 3).

Article Snippet: C2C12 cells grown to 30% confluence were transfected with 1 ng of vector containing MYOG shRNA or the scrambled vector using transfection reagent (Santa Cruz Biotechnology, CA, USA).

Techniques: Transfection, Incubation, Expressing, Quantitative RT-PCR, Knockdown, Gene Expression, Western Blot

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Pyruvate dehydrogenase kinase 4 promotes ubiquitin–proteasome system‐dependent muscle atrophy

doi: 10.1002/jcsm.13100

Figure Lengend Snippet: Knockdown of PDK4 prevents glucocorticoids‐induced muscle atrophy in vitro. (A) Immunoblot analysis of indicated protein in C2C12 myotubes after being treated with dexamethasone (DEX) for 24 h with different concentrations of 0, 10, 50 and 100 μM. (Right) Quantification represents the levels of the indicated protein normalized to HSP90. (B) qPCR analysis of indicated genes in C2C12 myotubes transfected with sh Pdk4 or shControl and treated with or without dexamethasone (DEX) (100 μM) for 24 h. (C) Immunoblot analysis of indicated proteins in C2C12 myotubes transfected with sh Pdk4 or shControl and treated with or without dexamethasone (DEX) (100 μM) for 24 h. (Right) Quantification represents the levels of the indicated protein normalized to HSP90. (D, E) Representative image of MYOG and MYHC immunofluorescence of C2C12 myotubes transfected with sh Pdk4 or shControl and treated with or without dexamethasone (DEX) (100 μM) for 24 h. (Below) Quantification represents the levels of the MYOG + ve cell and (Below) fusion index. (F) The distribution of nuclei per myotubes were calculated at Day 6 differentiation. Yellow arrowheads number of nuclei in one myotube. Mean ± SEM. (D and E) Scale bar 50 μM. (A–C) n = 3. (D, E) n = 4; number of image capture sites 20 from each group. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Dunnett's multiple comparisons test was used for the analysed panel (A). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Tukey's multiple comparisons test was used for the analysed panels (B–F). ROI, region of interest. Red * = nonspecific band.

Article Snippet: Briefly, PDK4 recombinant protein (provided by Dr Nam‐Ho Jeoung) and human recombinant MYOG protein (#H00004656‐P01; Novus Biological) were mixed in 1X‐kinase‐buffer (#9802S; Cell Signaling) and incubate 30 min at 30°C in the presence of 500‐μM ATP (A2383; Sigma).

Techniques: Knockdown, In Vitro, Western Blot, Transfection, Immunofluorescence

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Pyruvate dehydrogenase kinase 4 promotes ubiquitin–proteasome system‐dependent muscle atrophy

doi: 10.1002/jcsm.13100

Figure Lengend Snippet: PDK4 is upregulated in human and mouse models of muscle atrophy. (A) Immunoblot analysis of indicated proteins in gastrocnemius anterior (GA) muscles from obese, corticotrophin releasing hormone (CRH)‐Tg mice, and Type 2 diabetes (T2D) human myotube. (Right) Quantification represents the levels of the indicated protein normalized to HSP90. (B) Immunoblot analysis of indicated proteins in human gluteus maximus muscles (GM) from a normal subject and steroid patients. (Right) Quantification represents the levels of the indicated protein normalized to ACTB. (C) Representative image of haematoxylin and eosin staining of myofibre cross section isolated from human gluteus maximus (hGM) muscles. A microscope with a ×20 objective was used to capture the images. (Right) the cross‐sectional diameter of human gluteus maximus (GM) muscles. (D) (GSE169571) RNA sequencing analysis of quadriceps muscles from saline and dexamethasone (DEX) treated mouse. (E) Immunoblot analysis of indicated proteins in VXY and FLAG‐tagged PDK4 overexpressing C2C12 myotubes stable cells. (Right) Quantification represents the levels of the indicated protein normalized to HSP90. (F) Representative image of MYOG immunofluorescence in VXY and PDK4‐flag expressing C2C12 myotubes stable cells. (Right) Quantification represents the levels of the MYOG + ve cell (Mean ± SEM). (C, F) Scale bar 50 μM. (A, D and E) n = 3. (F) n = 4; number of image capture sites 20 from each group. (B, C) n = 6 human per group. ** P < 0.01; *** P < 0.001; **** P < 0.0001. Unpaired t ‐test was used for all analyses.

Article Snippet: Briefly, PDK4 recombinant protein (provided by Dr Nam‐Ho Jeoung) and human recombinant MYOG protein (#H00004656‐P01; Novus Biological) were mixed in 1X‐kinase‐buffer (#9802S; Cell Signaling) and incubate 30 min at 30°C in the presence of 500‐μM ATP (A2383; Sigma).

Techniques: Western Blot, Muscles, Staining, Isolation, Microscopy, RNA Sequencing, Saline, Immunofluorescence, Expressing

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Pyruvate dehydrogenase kinase 4 promotes ubiquitin–proteasome system‐dependent muscle atrophy

doi: 10.1002/jcsm.13100

Figure Lengend Snippet: Ectopic overexpression of MYOG mutant protects against glucocorticoids‐induced muscle atrophy in human myotube and mice. (A) Immunoblots analysis of indicated proteins in human myotubes infected with adenovirus expressing mutant MYOGᴬᴬ (Ad MYOGᴬᴬ) or GFP (AdGFP). Myotubes were culture for 24 h in the presence or absence of dexamethasone (DEX) 100 μM. (Below) Quantification represents the levels of the indicated protein normalized to HSP90. (B) Representative image of MYHC immunofluorescence in human myotubes infected with adenovirus expressing mutant MYOGᴬᴬ (Ad MYOGᴬᴬ) or GFP (AdGFP) in the presence or absence of dexamethasone (DEX) 100 μM for 24 h. (Below) Quantification represents the levels of fusion index and the distribution of nuclei per myotube were calculated at Day 6 differentiation. (C) The distribution of nuclei per myotubes were calculated at Day 6 differentiation. Yellow arrowheads indicate the number of nuclei in one myotube. (D) Immunoblots analysis of indicated proteins in gastrocnemius anterior (GA) muscle. (Right) Quantification represents the levels of the indicated protein normalized to HSP90. (E) The ratio of the weight of gastrocnemius anterior (GA) to total body weight. (F) Representative image of haematoxylin and eosin staining of myofibre cross section from AdGFP or AdMYOGᴬᴬ injected in gastrocnemius anterior muscles from dexamethasone (DEX)‐treated mice. A microscope with a ×20 objective was used to capture the images. (Right) the cross‐sectional diameter of gastrocnemius anterior (GA) muscle. Mean ± SEM. (B) Scale bar 50 μM. (F) n = 5 for each group. (A, E) n = 3. (B, C) n = 4; number of image capture sites 20 from each group. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Unpaired t ‐test was used for analyzed for panel (E–F). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Tukey's multiple comparisons test was used for analysed panels (A–C). ROI = region of interest.

Article Snippet: Briefly, PDK4 recombinant protein (provided by Dr Nam‐Ho Jeoung) and human recombinant MYOG protein (#H00004656‐P01; Novus Biological) were mixed in 1X‐kinase‐buffer (#9802S; Cell Signaling) and incubate 30 min at 30°C in the presence of 500‐μM ATP (A2383; Sigma).

Techniques: Over Expression, Mutagenesis, Western Blot, Infection, Expressing, Immunofluorescence, Staining, Injection, Muscles, Microscopy