Journal: International Journal of Molecular Sciences

Article Title: All- Trans Retinoic Acid-Responsive LGR6 Is Transiently Expressed during Myogenic Differentiation and Is Required for Myoblast Differentiation and Fusion

doi: 10.3390/ijms24109035

Figure Lengend Snippet: Expression pattern and role of LGR6 during myoblast differentiation. ( A ) C2C12 myoblasts were differentiated into myotubes for 5 days. The expression level of Lgr6 mRNA was determined by qPCR. ( B ) Myoblasts were differentiated into myotubes for 24 h. The levels of mRNAs encoding LGR6 and myogenic regulatory factors were determined by qPCR. ( C ) Myoblasts were transfected with control siRNA (siControl) or Lgr6 siRNA (siLGR6 #1 and siLGR6 #2), and cells were harvested 0 h after the induction of differentiation. The Lgr6 mRNA levels were determined by qPCR. ( D ) Myoblasts were differentiated for 3 days after siRNA transfection. Fixed cells were fluorescently labeled using an anti-MyHC antibody and fluorescence-labeled secondary antibody (green). The nuclei were stained with DAPI (blue). Bars, 100 μm. ( E ) The differentiation and fusion indices were calculated. The percentage of MyHC-positive cells with one, two, or three more nuclei was determined. ( F ) Myoblasts were differentiated for 24 h after siRNA transfection, and cells were harvested. The levels of mRNAs for myogenic regulatory factors were determined by qPCR. ( G ) Myoblasts were differentiated into myotubes. LGR6 expression was analyzed by Western blotting. Arrow indicates the lower band. ( H ) LGR6 levels were normalized to β-actin levels. ( A , B , H ) The results are presented as the mean ± SD ( n = 3). Data were determined using one-way ANOVA and Tukey’s post hoc test. ( A , H ) Columns with different letters are significantly different at p < 0.05, whereas columns sharing the same letters are not significantly different. ( B ) Different letters with the same color on the lines indicate statistically significant differences ( p < 0.05). ( C , E , F ) The results are presented as the mean ± SD ( n = 3). Data were determined using one-way ANOVA and Dunnet’s post hoc test. * p < 0.05 vs. siControl.

Article Snippet: Murine C2C12 myoblasts (European Collection of Authenticated Cell Cultures, Salisbury, UK) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and antibiotics (growth medium) and differentiated into myotubes in Dulbecco’s modified Eagle’s medium supplemented with 2% horse serum and antibiotics (differentiation medium), as described previously [ ].

Techniques: Expressing, Transfection, Control, Labeling, Fluorescence, Staining, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: All- Trans Retinoic Acid-Responsive LGR6 Is Transiently Expressed during Myogenic Differentiation and Is Required for Myoblast Differentiation and Fusion

doi: 10.3390/ijms24109035

Figure Lengend Snippet: LGR6 expression during myoblast differentiation as an ATRA-responsive gene. ( A ) C2C12 myoblasts were differentiated in the presence or absence of ATRA. The Lgr6 mRNA levels were determined by qPCR. ( B ) Myoblasts were cultured with ATRA in the presence of AGN193109. The Lgr6 mRNA levels were determined by qPCR. ( C ) C2C12 myoblasts were cultured with ATRA, AM580, BMS961, AM580, and BMS961. The Lgr6 mRNA levels were determined by qPCR. ( D ) Myoblasts were cultured with ATRA in the presence or absence of Ro41-5253 and/or LY2955303. The Lgr6 mRNA levels were determined by qPCR. ( E ) Myoblasts were transfected with control siRNA (siControl) or Lgr6 siRNA (siLGR6#1), and cells were harvested immediately after the induction of differentiation. The Lgr6 mRNA levels were determined by qPCR. ( F ) After siRNA transfection, myoblasts were differentiated in the presence or absence of ATRA for 3 days. Fixed cells were immunofluorescently labeled using an anti-MyHC antibody (green), and the nuclei were stained with DAPI (blue). Bars, 100 μm. ( G ) The differentiation and fusion indices were calculated. ( A – D , G ) The results are presented as the mean ± SD ( n = 3). Data were determined using two-way ANOVA and Tukey’s post hoc test. Columns with different letters are significantly different at p < 0.05, whereas columns sharing the same letters are not significantly different. ( E ) The results are presented as the mean ± SD ( n = 3). Data were determined using Student’s t -test. * p < 0.05 vs. siControl.

Article Snippet: Murine C2C12 myoblasts (European Collection of Authenticated Cell Cultures, Salisbury, UK) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and antibiotics (growth medium) and differentiated into myotubes in Dulbecco’s modified Eagle’s medium supplemented with 2% horse serum and antibiotics (differentiation medium), as described previously [ ].

Techniques: Expressing, Cell Culture, Transfection, Control, Labeling, Staining

Journal: International Journal of Molecular Sciences

Article Title: All- Trans Retinoic Acid-Responsive LGR6 Is Transiently Expressed during Myogenic Differentiation and Is Required for Myoblast Differentiation and Fusion

doi: 10.3390/ijms24109035

Figure Lengend Snippet: Involvement of the ubiquitin–proteasome system in LGR6 expression. ( A ) C2C12 myoblasts were transfected with a mock vector and murine and human LGR6 expression vectors, followed by culture in the presence or absence of MG132. LGR6 levels were analyzed by Western blotting. ( B ) Myoblasts were transfected with a mock vector and a human LGR6 expression vector, followed by further culture in the presence or absence of MG132. The LGR6 levels were analyzed by Western blotting. Myc- and His-tagged LGR6 (LGR6-Myc/His) was pulled down using Ni-Sepharose resin, and Ni-Sepharose-bound proteins were analyzed by Western blotting using anti-Myc and anti-ubiquitin antibodies. ( C ) Myoblasts were transfected with a human LGR6 expression vector, followed by further transfection with control siRNA or Znrf3 siRNAs (siZNRF3#1 and siZNRF3#2). As a negative control for exogenous LGR6, myoblasts were transfected with a mock vector. The Lgr6 mRNA levels were determined by qPCR. Data were determined by Student’s t -test. * p < 0.05 vs. siControl. ( D ) Exogenous LGR6 levels were analyzed by Western blotting and normalized to β-actin levels. The results are presented as the mean ± SD ( n = 3). Data were determined using one-way ANOVA and Dunnet’s post hoc test. * p < 0.05 vs. siControl. ( E ) Deduced positions of lysine residues (yellow stars: K598, K679, and K861) in the intracellular region of human LGR6. ( F ) Myoblasts were transfected with a mock vector and wild-type or mutant-type human LGR6 expression vectors, followed by further culture in the presence or absence of MG132. LGR6 expression was analyzed by Western blotting using an anti-Myc antibody.

Article Snippet: Murine C2C12 myoblasts (European Collection of Authenticated Cell Cultures, Salisbury, UK) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and antibiotics (growth medium) and differentiated into myotubes in Dulbecco’s modified Eagle’s medium supplemented with 2% horse serum and antibiotics (differentiation medium), as described previously [ ].

Techniques: Ubiquitin Proteomics, Expressing, Transfection, Plasmid Preparation, Western Blot, Control, Negative Control, Mutagenesis

Journal: International Journal of Molecular Sciences

Article Title: All- Trans Retinoic Acid-Responsive LGR6 Is Transiently Expressed during Myogenic Differentiation and Is Required for Myoblast Differentiation and Fusion

doi: 10.3390/ijms24109035

Figure Lengend Snippet: Involvement of LGR6 in Wnt/β-catenin signaling. ( A ) C2C12 myoblasts were transfected with a TCF reporter expression vector, followed by further incubation in the presence or absence of RSPO2. TCF activity was determined. ( B ) Myoblasts were transfected with a TCF reporter expression vector and a Wnt3a expression vector, followed by further incubation in the presence or absence of RSPO2. TCF activity was determined. ( C ) Myoblasts were transfected with a TCF reporter expression vector and a Wnt3a expression vector and further transfected with control siRNA (siControl) or Lgr6 siRNAs (siLGR6 #1 and siLGR6 #2). Cells were harvested immediately after the induction of differentiation. The Lgr6 mRNA levels were determined by qPCR. ( D ) After siRNA transfection, myoblast differentiation was induced in the presence or absence of RSPO2. TCF activity was determined. ( A , B , D ) The results are presented as the mean ± SD ( n = 3). Data were determined using two-way ANOVA and Tukey’s post hoc test. Columns with different letters are significantly different at p < 0.05, whereas columns sharing the same letters are not significantly different. ( C ) The results are presented as the mean ± SD ( n = 3). Data were determined using Student’s t -test. * p < 0.05 vs. siControl.

Article Snippet: Murine C2C12 myoblasts (European Collection of Authenticated Cell Cultures, Salisbury, UK) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and antibiotics (growth medium) and differentiated into myotubes in Dulbecco’s modified Eagle’s medium supplemented with 2% horse serum and antibiotics (differentiation medium), as described previously [ ].

Techniques: Transfection, Expressing, Plasmid Preparation, Incubation, Activity Assay, Control

Journal: Biomolecules

Article Title: Decapeptide from Potato Hydrolysate Induces Myogenic Differentiation and Ameliorates High Glucose-Associated Modulations in Protein Synthesis and Mitochondrial Biogenesis in C2C12 Cells

doi: 10.3390/biom12040565

Figure Lengend Snippet: DI-10 activates ERK, Akt, mTOR, and FOXO3a proteins and facilitates protein synthesis in myoblast cells. Dose-dependent activation of ERK, Akt, mTOR, and FOXO3a was observed in myoblast cells after DI-10. ( A ) Western blotting show phosphorylation of ERK, Akt, mTOR, and FOXO3A in C2C12 cells after DI-10 treatment. ( B ) Graph showing densitometric analysis. *** p < 0.001, and ** p < 0.01 compared to control group.

Article Snippet: C2C12 murine myoblast cells was purchased from Bioresource Collection and Research Center (BCRC, Taiwan) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS) containing antibiotics streptomycin (100 μg/mL) and penicillin (100 U/mL) in a 10 cm culture plate.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Control

Journal: Biomolecules

Article Title: Decapeptide from Potato Hydrolysate Induces Myogenic Differentiation and Ameliorates High Glucose-Associated Modulations in Protein Synthesis and Mitochondrial Biogenesis in C2C12 Cells

doi: 10.3390/biom12040565

Figure Lengend Snippet: DI-10 improves the viability of C2C12 myoblast cells under high glucose stress. ( A ) C2C12 cells were treated with different concentrations of D-glucose (5, 15, and 30 mM) and the cell viability was determined with MTT assay. ( B ) C2C12 cells were treated with 30 mM HG followed by different concentrations of DI-10. *** p < 0.001 compared to control groups and ### p < 0.001 compared to the high glucose.

Article Snippet: C2C12 murine myoblast cells was purchased from Bioresource Collection and Research Center (BCRC, Taiwan) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS) containing antibiotics streptomycin (100 μg/mL) and penicillin (100 U/mL) in a 10 cm culture plate.

Techniques: MTT Assay, Control

Journal: Biomolecules

Article Title: Decapeptide from Potato Hydrolysate Induces Myogenic Differentiation and Ameliorates High Glucose-Associated Modulations in Protein Synthesis and Mitochondrial Biogenesis in C2C12 Cells

doi: 10.3390/biom12040565

Figure Lengend Snippet: Decapeptide DI-10 promotes myoblast differentiation under high-glucose conditions. Effect of decapeptide DI-10 on myogenic differentiation under hyperglycemic conditions was determined by ( A ) microscopic observation of cell morphology. Photomicrographs were obtained using an Olympus ® CKX53 microscope (100×) ( B ) MyHC expression in C2C12 cells under HG conditions. ### p < 0.001 compared to the high glucose.

Article Snippet: C2C12 murine myoblast cells was purchased from Bioresource Collection and Research Center (BCRC, Taiwan) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS) containing antibiotics streptomycin (100 μg/mL) and penicillin (100 U/mL) in a 10 cm culture plate.

Techniques: Microscopy, Expressing

Journal: Biomolecules

Article Title: Decapeptide from Potato Hydrolysate Induces Myogenic Differentiation and Ameliorates High Glucose-Associated Modulations in Protein Synthesis and Mitochondrial Biogenesis in C2C12 Cells

doi: 10.3390/biom12040565

Figure Lengend Snippet: Effect of DI-10 on Akt, mTOR, and AMPK phosphorylation under hyperglycemic conditions in C2C12 cells. ( A ) Immuno blot showing phosphorylation of Akt, mTOR, and AMPK proteins involved in protein synthesis. ( B ) Graph showing densitometric analysis. ** p < 0.01 and * p < 0.05 compared to control group and # p < 0.05 and ### p < 0.001 compared to the high glucose.

Article Snippet: C2C12 murine myoblast cells was purchased from Bioresource Collection and Research Center (BCRC, Taiwan) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS) containing antibiotics streptomycin (100 μg/mL) and penicillin (100 U/mL) in a 10 cm culture plate.

Techniques: Phospho-proteomics, Control

Journal: Biomolecules

Article Title: Decapeptide from Potato Hydrolysate Induces Myogenic Differentiation and Ameliorates High Glucose-Associated Modulations in Protein Synthesis and Mitochondrial Biogenesis in C2C12 Cells

doi: 10.3390/biom12040565

Figure Lengend Snippet: Bioactive peptides enhance myogenic differentiation and skeletal muscle protein synthesis under high-glucose conditions in C2C12 myoblast cells. ↑ increase, ↓ decrease.

Article Snippet: C2C12 murine myoblast cells was purchased from Bioresource Collection and Research Center (BCRC, Taiwan) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) with 10% Fetal Bovine Serum (FBS) containing antibiotics streptomycin (100 μg/mL) and penicillin (100 U/mL) in a 10 cm culture plate.

Techniques:

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: a Cytotoxic effects of teritiary butyl hydroperoxide (t-BHP) on C2C12 muscle cells. *P < 0.01 versus control cells without any treatment. b Dose dependent protective effect of treatment with Celastrus paniculatus seed extract (CPSE) on t-BHP induced cytotoxicity in C2C12 cells, the cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. *P < 0.01 versus control cells and 250 µM t-BHP treated group. c Cytotoxic effects of t-BHP on C2C12 muscle cell analyzed by lactate dehydrogenase (LDH) leakage assay. #P < 0.01 versus control group *P < 0.01 versus control cells. d Protective effect of CPSE pretreatment on t-BHP (250 μm) induced cytotoxicity by LDH leakage assay. *P < 0.01 versus control cells and 250 µM t-BHP treated group. #P < 0.01 versus control group. e Effects of CPSE treated on t-BHP induced morphological alterations in C2C12 muscle cells by phase-contrast microscopy. The data are represented as mean ± SD of three independent experiments

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Control, MTT Assay, Microscopy

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: a Estimation of mitochondrial membrane potential in C2C12 muscle cells with pre-treatment of CPSE on t-BHP challenge. The fluorescence intensity was determined using a spectrofluorimeter. b The membrane potential was monitored by fluorescent microscope (Olympus) Control cells without any treatment (a), 50 µg/ml CPSE treatment for 2 h (b), 250 µM t-BHP treatment for 24 h (c), and cells pre-treated with 50 µg/ml CPSE for 2 h then treated with 250 µM t-BHP for 24 h (d). The data are represented as mean ± SD of three independent experiments. #P < 0.01 versus control group, *P < 0.01 versus control cells and *P < 0.01 versus 250 µM t-BHP treated group

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Membrane, Fluorescence, Microscopy, Control

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: a Estimation of intracellular ROS production using 2′,7′,DCFH2DA in C2C12 muscle cells pre-treated with CPSE on t-BHP challenge by spectrofluorimeter. b The membrane potential was monitored by fluorescence microscope (Olympus). Control cells without any treatment (a), 50 µg/ml CPSE treatment for 2 h (b), 250 µM t-BHP treatment for 24 h (c), and cells pre-treated with 50 µg/ml CPSE for 2 h then treated with 250 µM t-BHP for 24 h (d). The data are represented as mean ± SD of three independent experiments. #P < 0.01 versus control group, *P < 0.01 versus control cells and *P < 0.01 versus 250 µM t-BHP treated group

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Membrane, Fluorescence, Microscopy, Control

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: Estimation of lipid peroxidation products by thiobarbituric acid (TBARS) assay in C2C12 muscle cells with pre-treatment of CPSE on t-BHP challenge. The data are represented as mean ± SD of three independent experiments. #P < 0.01 versus control group, *P < 0.01 versus control cells and 250 µM t-BHP treated group

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: TBARS Assay, Control

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: Pre-treatment of CPSE on restoration of SOD and catalase enzyme activities in C2C12 muscle cells challenged with t-BHP. The data are represented as mean ± SD of three independent experiments. #P < 0.01 versus control group, *P < 0.01 versus control cells and *P < 0.01 versus 250 µM t-BHP treated group

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Control

Journal: Cytotechnology

Article Title: Celastrus paniculatus Willd. mitigates t-BHP induced oxidative and apoptotic damage in C2C12 murine muscle cells

doi: 10.1007/s10616-014-9733-0

Figure Lengend Snippet: Protective effect of CPSE on DNA damage induced by t-BHP in C2C12 cells. Control cells without any treatment (a), Cells with 250 µM t-BHP treatment for 24 h (b), C2C12 cells were pre-treated with CPSE for 2 h at 50 µg/assay and treated with 250 μM t-BHP (duration: 24 h) (c). Tail length (µm) Bars. Inhibitory effect of CPSE on DNA damage diamond (d)

Article Snippet: C2C12 cell differentiation and treatments Murine C2C12 myoblasts were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Control

Journal: Annals of biomedical engineering

Article Title: Modeling the effect of TNF-α upon drug induced toxicity in human, tissue-engineered myobundles.

doi: 10.1007/s10439-019-02263-8

Figure Lengend Snippet: TNF-α decreases total cellular protein and increases proliferation of differentiating C2C12 and human myoblasts in 2D cultures. TNF-α (0–10,000U/mL) was added daily at the onset of differentiation to (A-C) C2C12 myoblasts for 3 days and (D-F) human myoblasts differentiated for 5 days. A) C2C12 total protein content measured by BCA assay and normalized by mass of DNA for control and TNF-α dose. B) C2C12 DNA mass of myotubes per plate. C) Percent of C2C12 myoblasts proliferating measured by percent of total nuclei staining positive for EdU. Myoblasts were incubated for 2 hours with EdU components after 24 hours of differentiation and TNF-α dosage. D) Normalized protein content and E) DNA mass per plate of hSkM myoblasts on day 3 of differentiation and TNF-α dose. F) Percent of hSkM myoblasts proliferating measured by nuclei staining positive for EdU. Proliferating myoblasts were incubated for 2 hours with EdU components after 24 hours of differentiation and TNF-α dose. Data are mean±SEM for duplicate wells from each of 4 donors. Significance was determined using one-way ANOVA with Bonferroni post-hoc corrections.

Article Snippet: C2C12 myoblast culture The murine C2C12 cell line myoblasts were cultured in C2C12 growth media (C2C12 GM) containing high glucose (HG;4.5g/L D-glucose) DMEM, 8% fetal bovine serum, 8% bovine calf serum (Hyclone), 0.5% chick embryo extract (Accurate Chemical), and 1x antibiotic-antimycotic.

Techniques: BIA-KA, Staining, Incubation

Journal: Annals of biomedical engineering

Article Title: Modeling the effect of TNF-α upon drug induced toxicity in human, tissue-engineered myobundles.

doi: 10.1007/s10439-019-02263-8

Figure Lengend Snippet: TNF-α inhibits MHC expression in both C2C12 and human differentiating myoblasts in 2D culture. Immunofluorescent staining for MHC (green) and Hoechst staining for nuclei (blue) revealed the extent of myoblast differentiation at varying concentrations of TNF-α (0-10,000U/mL) after 5 days of daily exposure. There was a noticeable decrease in the expression of MHC in representative images of (A-D) C2C12 differentiating myoblasts cultured with A) vehicle control, B) 100U/mL, C) 1,000U/mL, D) 10,000U/mL of TNF-α. A similar decrease was also seen in representative images of (E-H) hSkM differentiating myoblasts cultured with E) vehicle control, F) 100U/mL, G) 1,000U/mL, H) 10,000U/mL of TNF-α. Scale bar=200μm. The percent of nuclei in MHC-positive fibers and the percent of the total image staining MHC-positive indicated the degree of maturation in differentiating (I,J,M) C2C12 and (K,L,N) hSkM myoblasts after exposure to different TNF-α for 5 days. All measurements were obtained from immunofluorescent images. I) Percent of C2C12 nuclei in MHC-positive fibers. J) Percent area in the field of view stained positive for MHC in C2C12 cultures. K) Percent of nuclei in MHC-positive fibers in hSkM cultures separated by donor. L) Percent field of view area stained positive for MHC in hSkM cultures separated by donor. M) Number of C2C12 nuclei per fiber. N) Number of hSKM nuclei per fiber. Data are mean±SEM for duplicate wells with 5 fields of view per well from 4 donors for panels I,J,M and N and mean±SEM for duplicate wells with 5 fields of view per well for panels K and L. Significance was determined using one-way or two-way ANOVA with Bonferroni post-hoc corrections.

Article Snippet: C2C12 myoblast culture The murine C2C12 cell line myoblasts were cultured in C2C12 growth media (C2C12 GM) containing high glucose (HG;4.5g/L D-glucose) DMEM, 8% fetal bovine serum, 8% bovine calf serum (Hyclone), 0.5% chick embryo extract (Accurate Chemical), and 1x antibiotic-antimycotic.

Techniques: Expressing, Staining, Cell Culture

Journal: Annals of biomedical engineering

Article Title: Modeling the effect of TNF-α upon drug induced toxicity in human, tissue-engineered myobundles.

doi: 10.1007/s10439-019-02263-8

Figure Lengend Snippet: Myotubes exposed to TNF-α exhibit less fusion in 2D culture. The degree to which myoblasts fuse into multinucleated myotubes is another indicator of skeletal muscle maturation. All (A,B) C2C12 and (C,D) hSkM measurements were obtained by quantifying immunofluorescent images. Data are mean±SEM for duplicate wells from 4 donors. 5 randomly chosen images were analyzed per well. A) Average C2C12 myotube diameter. B) Average C2C12 myotube length. C) Average hSkM myotube diameter. D) Average hSkM myotube length.

Article Snippet: C2C12 myoblast culture The murine C2C12 cell line myoblasts were cultured in C2C12 growth media (C2C12 GM) containing high glucose (HG;4.5g/L D-glucose) DMEM, 8% fetal bovine serum, 8% bovine calf serum (Hyclone), 0.5% chick embryo extract (Accurate Chemical), and 1x antibiotic-antimycotic.

Techniques: