Journal: Animal Cells and Systems

Article Title: Phosphoinositide 3-kinase regulates myogenin expression at both the transcriptional and post-transcriptional level during myogenesis

doi: 10.1080/19768354.2010.496541

Figure Lengend Snippet: Figure 2. PI3-kinase regulates myogenin expression at both the post-transcriptional and transcriptional level. (A) At the time of inducing differentiation (0 h), the myoblasts were exposed to the indicated concentrations of LY294002 and cultured for 24 h. The cells were assayed for myogenin expression by Western blotting and qRT-PCR analysis. b-actin was used as a loading control for both experiments. (B) At 40 h after inducing differentiation, the cells were treated with () or without () 20 mM of LY294002 and cultured for the indicated times. The protein and mRNA levels of myogenin and GAPDH were determined using Western blotting and qRT-PCR analysis. GAPDH was used as a loading control. Results are representative of either three or four independent experiments.

Article Snippet: The antibodies for myogenin, b-actin and glyceraldehyde 3-phosphate (GAPDH) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Cell Culture, Western Blot, Quantitative RT-PCR, Control

Journal: Animal Cells and Systems

Article Title: Phosphoinositide 3-kinase regulates myogenin expression at both the transcriptional and post-transcriptional level during myogenesis

doi: 10.1080/19768354.2010.496541

Figure Lengend Snippet: Figure 3. Measurement of myogenin protein and mRNA stability. (A) At 40 h after inducing differentiation, the cells were treated with () or without () 10 ng/ml of cyclohex- imide and cultured for the indicated times. The protein and mRNA levels of myogenin and GAPDH were determined by Western blotting and qRT-PCR analysis. GAPDH was used as a loading control. (B) To determine mRNA stability, the cells were incubated with 2 mg/ml of actinomycin D in the presence or absence of LY294002 (20 mM) at 40 h after inducing differentiation and cultured for the indicated times. The mRNA level of myogenin was determined by qRT- PCR analysis. GAPDH was used as a loading control. Results are representative of either three or four independent experiments.

Article Snippet: The antibodies for myogenin, b-actin and glyceraldehyde 3-phosphate (GAPDH) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Cell Culture, Western Blot, Quantitative RT-PCR, Control, Incubation

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: Journal of Biomedical Science

Article Title: Hypoxia and therapeutic treatment of EV-A71 with an immune modulator TLR7 agonist in a new immunocompetent mouse model

doi: 10.1186/s12929-019-0585-y

Figure Lengend Snippet: A “white-jade” muscle phenotype in EV-A71-infected mouse strain 129. a Left panel : We coined a “white-jade” phenotype to describe the pathological changes in the color of the hindlimb muscle in EV-A71-infected mice (indicated by arrow). This phenotype showed characteristic muscle with locally whitened color in appearance and hardened tissue mass. In the saline control, tendons (not muscle) can be seen in white color. Right panel : An anatomical sketch of the hindlimb muscle. GM: gluteal muscle, QF: quadratus femoris muscle, BF: Biceps femoris muscle. b Both VP1 protein (upper) and myogenin protein (lower) were detected by IHC staining in the muscles, indicating EV-A71 infection, muscle injury and regeneration. The magnification is 200X. c Striking expression of HIF1A (hypoxia inducible factor 1-α) protein (brown color) was detected by IHC staining in the white-jaded tissue, but not in the non-white-jaded tissue from the same mouse. d Specific association between HIF1A expression and white-jaded muscle

Article Snippet: Slides were washed with PBS containing 0.1% Tween 20 (PBST), followed by incubation with specific antibodies, including rabbit anti-VP1 polyclonal antibody (PB7631, Abnova, Taiwan), rabbit anti-HIF1A polyclonal antibody (NB100–479, Novus), rabbit anti-VEGFA polyclonal antibody (ab46154, Abcam), rabbit anti-myogenin monoclonal antibody (GTX 63352, Genetex, Taiwan), as well as antibodies specific for the immune system, CD3 (MA1–90582, Thermo), CD19 (PAB19567, Abnova), CD68 (ab955, Abcam), and CD163 (bs-2527, Bioss), respectively.

Techniques: Infection, Saline, Control, Immunohistochemistry, Muscles, Expressing

Journal: Journal of Biomedical Science

Article Title: Hypoxia and therapeutic treatment of EV-A71 with an immune modulator TLR7 agonist in a new immunocompetent mouse model

doi: 10.1186/s12929-019-0585-y

Figure Lengend Snippet: Hypoxia in EV-A71-infected muscle was rescued by timely treatment with IFNαA. a A schematic diagram of mouse IFNαA treatment after EV-A71 infection. Six different experimental groups of mice received mIFNαA (300 IU/g of mouse) at different dpi and frequencies of treatments. Therapeutic efficacies were assayed by survival rates. In some cases, infected and treated mice ( n = 12) were monitored for up to 1.5 years. b Clinical scores were significantly reduced only in Exp. I (green) and Exp. II (red), when treated at dpi 1, or dpi 1, 2, 3. c Survival rates of Exp. I (green) (100%) and Exp. II (red) (88%) were significantly higher than that of the control group with no IFNαA treatment. d No VP1-positive signal was detected in the muscle by IHC at 25 dpi and 41 dpi in mice recovered from IFNαA treatment in Exp. 1. e The myogenin protein (black arrowhead) was expressed in recovered muscle via IHC staining, indicating muscle regeneration after injury. f HIF1A and VEGFA proteins were both expressed in newly recovered muscle (dpi = 25), but not in the fully recovered muscle (dpi = 41)

Article Snippet: Slides were washed with PBS containing 0.1% Tween 20 (PBST), followed by incubation with specific antibodies, including rabbit anti-VP1 polyclonal antibody (PB7631, Abnova, Taiwan), rabbit anti-HIF1A polyclonal antibody (NB100–479, Novus), rabbit anti-VEGFA polyclonal antibody (ab46154, Abcam), rabbit anti-myogenin monoclonal antibody (GTX 63352, Genetex, Taiwan), as well as antibodies specific for the immune system, CD3 (MA1–90582, Thermo), CD19 (PAB19567, Abnova), CD68 (ab955, Abcam), and CD163 (bs-2527, Bioss), respectively.

Techniques: Infection, Control, Immunohistochemistry

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: (A, B) Myogenin expression in iPSC-derived myogenic cells (day 3) evaluated by intracellular flow cytometry. (A, B) Representative flow diagrams and (B) quantification. (C) qPCR analysis of MyoG and MEF2C sequential expression. (D) Immunoblotting assay of SMN with iPSC-derived myogenic cells (day 6). (E) Cell number during myogenic conversion. (F) Change in relative mitochondrial DNA copy number in iPSC-derived myogenic cells. (G) Spare respiratory capacity was calculated using the data in . Data were obtained from 1.0 × 10 4 cells. (H) Mitochondrial membrane potential (∆ψm) of iPSC-derived myogenic cells (day 6). (I) Level of reactive oxygen species in iPSC-derived myogenic cells (day 3) (relative to the MFI of B7-M or SMA-MOE). (J) Cleaved caspase-3–positive apoptotic cells in SMN-down-regulated clones (day 3) analyzed by intracellular flow cytometry. (K) Number of iPSC-derived myogenic cells (day 3). (L, M) Expression of Smn in C2C12SC and C2C12siSmn (day 6) at the mRNA (L) and protein level (M). (N, O) Expression of COX1 protein in C2C12 cells (day 6) (N) and quantification (O). Value for day 0 is set as 1, and the relative values are shown. Error bars indicate means ± SD. (I, J, K) αToc, α-tocopherol (100 nM). (K) Statistical analysis by one-way ANOVA with multiple comparisons. (F, G, H, I, J) Statistical analysis by a t test. Each dot represents a biologically independent sample.r.

Article Snippet: Then, the cells were labeled with antibodies against cleaved caspase-3 (1:20; #9602S; CST) or MyoG (1:20; #NBP2-33056; Novus Biologicals).

Techniques: Expressing, Derivative Assay, Flow Cytometry, Western Blot, Membrane, Clone Assay

Journal:

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

doi:

Figure Lengend Snippet: 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:

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

doi:

Figure Lengend Snippet: 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:

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

doi:

Figure Lengend Snippet: 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 rhabdomyosarcomas (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:

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

doi:

Figure Lengend Snippet: Summary of Anti-Myogenin Staining in Embryonal and Alveolar Rhabdomyosarcomas

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

Techniques: Staining

Journal:

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

doi:

Figure Lengend Snippet: Analysis of an Additional Subset of Rhabdomyosarcomas for Quantitative Differences in Expression of Myogenin by Western Blot Analysis and Correlation with Immunohistochemical Staining Data

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

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal:

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

doi:

Figure Lengend Snippet: Immunohistochemical Staining for Myogenin in Embryonal and Alveolar Rhabdomyosarcomas

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

Techniques: Immunohistochemical staining, Staining

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