Journal: Cancer Cell International

Article Title: In vitro myotoxic effects of bupivacaine on rhabdomyosarcoma cells, immortalized and primary muscle cells

doi: 10.1186/s12935-015-0229-6

Figure Lengend Snippet: Microscopic images of a C2C12 cells; b primary muscle cells and c rhabdomyosarcoma cells.

Article Snippet: Rhabdomyosarcoma cells (CLS-Cell Lines Service, Eppelheim, Germany), immortalized C2C12 (CLS-Cell Lines Service, Eppelheim, Germany) muscle cells and primary muscle cells were used for the study.

Techniques:

Journal: Cancer Cell International

Article Title: In vitro myotoxic effects of bupivacaine on rhabdomyosarcoma cells, immortalized and primary muscle cells

doi: 10.1186/s12935-015-0229-6

Figure Lengend Snippet: Calculated bupivacaine concentration (ppm) with 50% cell survival (IC50)

Article Snippet: Rhabdomyosarcoma cells (CLS-Cell Lines Service, Eppelheim, Germany), immortalized C2C12 (CLS-Cell Lines Service, Eppelheim, Germany) muscle cells and primary muscle cells were used for the study.

Techniques: Concentration Assay

Journal: Cancer Cell International

Article Title: In vitro myotoxic effects of bupivacaine on rhabdomyosarcoma cells, immortalized and primary muscle cells

doi: 10.1186/s12935-015-0229-6

Figure Lengend Snippet: Combined calculated IC50 bupivacaine concentrations (ppm) differentiating only cell type

Article Snippet: Rhabdomyosarcoma cells (CLS-Cell Lines Service, Eppelheim, Germany), immortalized C2C12 (CLS-Cell Lines Service, Eppelheim, Germany) muscle cells and primary muscle cells were used for the study.

Techniques:

Journal: Toxics

Article Title: Selenium Nanoparticles Attenuate Cobalt Nanoparticle-Induced Skeletal Muscle Injury: A Study Based on Myoblasts and Zebrafish

doi: 10.3390/toxics12020130

Figure Lengend Snippet: Cellular toxicity induced by cobalt and selenium nanoparticles. ( A ) Cells were treated with 5 μg/mL FITC-labeled CoNPs and SeNPs for 2 h. Confocal microscopy revealed fluorescent signals into the cells. Yellow arrows indicating cells that are filled with green fluorescence, while the culture medium contains free FITC-labeled CoNPs and SeNPs. ( B – D ) Viability of C2C12 cells treated with 5–80 μg/mL CoNPs, SeNPs, and mixing of CoNPs and SeNPs were determined by CCK-8. Data are presented as mean ± standard deviation of three identical experiments conducted in triplicate. * Statistically significant difference compared with the controls ( p < 0.05 for each).

Article Snippet: According to the manufacturer’s instructions, the levels of MDA (malondialdehyde), GSH (glutathione), and SOD (superoxide dismutase) in C2C12 cells were measured using a GSH ELISA Kit (Elabscience, Wuhan, China), MDA Content Detection Kit (Beyotime, Nantong, China), and SOD Activity Detection Kit (Beyotime, Nantong, China).

Techniques: Labeling, Confocal Microscopy, Fluorescence, CCK-8 Assay, Standard Deviation

Journal: Toxics

Article Title: Selenium Nanoparticles Attenuate Cobalt Nanoparticle-Induced Skeletal Muscle Injury: A Study Based on Myoblasts and Zebrafish

doi: 10.3390/toxics12020130

Figure Lengend Snippet: Protective effects of low-dose SeNPs on CoNP-induced oxidative stress in muscle cells. ( A ) C2C12 cells were exposed to control, 20 μg/mL CoNPs, 500 µM H2O2, 5 μg/mL SeNPs with or without H2O2, and mixing of Co and Se NPs. Oxidative stress condition induced by H2O2 treatment for 4 h. Intracellular ROS production was analyzed by H2DCFDA staining. ( B – E ) MDA, GSH, and SOD levels were determined in C2C12 cells from control, 20 μg/mL CoNPs, 500 µM H2O2, 5 μg/mL SeNPs with or without H2O2, and mixing of Co and Se NP-treated groups. Data are displayed as mean ± SD; * means p < 0.05 between two indicated groups (n = 10).

Article Snippet: According to the manufacturer’s instructions, the levels of MDA (malondialdehyde), GSH (glutathione), and SOD (superoxide dismutase) in C2C12 cells were measured using a GSH ELISA Kit (Elabscience, Wuhan, China), MDA Content Detection Kit (Beyotime, Nantong, China), and SOD Activity Detection Kit (Beyotime, Nantong, China).

Techniques: Control, Staining

Journal: Toxics

Article Title: Selenium Nanoparticles Attenuate Cobalt Nanoparticle-Induced Skeletal Muscle Injury: A Study Based on Myoblasts and Zebrafish

doi: 10.3390/toxics12020130

Figure Lengend Snippet: CoNP-induced apoptosis was inhibited by SeNPs in muscle cells. ( A , B ) The expression of caspase–3 and cleaved caspase–3 in C2C12 cells treated with 5 μg/mL SeNPs or 20 μg/mL CoNPs with or without SeNPs was analyzed by Western blotting. GAPDH was used as internal reference. Relative expression levels are shown in the graph. ( C , D ) The degree of apoptosis of cells subjected to different treatments was determined by annexin V-FITC/PI double-staining flow cytometry. The percentage of apoptotic cells was counted and compared between the different groups. Data are displayed as mean ± SD; * means p < 0.05 between two indicated groups (n = 3).

Article Snippet: According to the manufacturer’s instructions, the levels of MDA (malondialdehyde), GSH (glutathione), and SOD (superoxide dismutase) in C2C12 cells were measured using a GSH ELISA Kit (Elabscience, Wuhan, China), MDA Content Detection Kit (Beyotime, Nantong, China), and SOD Activity Detection Kit (Beyotime, Nantong, China).

Techniques: Expressing, Western Blot, Double Staining, Flow Cytometry

Journal: Toxics

Article Title: Selenium Nanoparticles Attenuate Cobalt Nanoparticle-Induced Skeletal Muscle Injury: A Study Based on Myoblasts and Zebrafish

doi: 10.3390/toxics12020130

Figure Lengend Snippet: Promotive effects of SeNPs on CoNP-induced inhibition of myogenic differentiation. ( A ) Immunofluorescence staining in SeNPs, CoNPs, and Co Se NP-treated C2C12 cells was analyzed at 4 days of differentiation to detect the effect of different nanoparticles on myogenic differentiation. Nuclei (DAPI, blue), F-actin (Green), and α-SMA (Red) were labeled; scale bar = 25 µm. ( B ) Quantification of α-SMA fluorescence intensity in ( A ) was performed. Data are displayed as mean ± SD. * means p < 0.05 between two indicated groups (n = 6). ( C – E ) mRNA expression of myogenic markers, MyoD, myogenin, and Myf5, in C2C12 cells treated with 5 μg/mL SeNPs, 20 μg/mL CoNPs with or without SeNPs was analyzed by qPCR. The control group was set to 1.0. Data are displayed as mean ± SD. * means p < 0.05 between two indicated groups (n = 6).

Article Snippet: According to the manufacturer’s instructions, the levels of MDA (malondialdehyde), GSH (glutathione), and SOD (superoxide dismutase) in C2C12 cells were measured using a GSH ELISA Kit (Elabscience, Wuhan, China), MDA Content Detection Kit (Beyotime, Nantong, China), and SOD Activity Detection Kit (Beyotime, Nantong, China).

Techniques: Inhibition, Immunofluorescence, Staining, Labeling, Fluorescence, Expressing, Control

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: Differentiation of C2C12 wild-type and Gne knockout clones. ( A ) Micrographs of differentiated C2C12 wild-type (WT) and Gne KO clone #24 and clone #26. ( B ) Immunofluorescent staining of Myh on day 7 of differentiation in C2C12 WT and Gne KO clones. ( C ) Quantification of the Myh signal norm to DAPI. Myh: myosin heavy chain I. α-ms Alexa647: negative control (secondary antibody only). Micrographs and immunofluorescent staining show representative pictures. Scale bar in ( A ) 200 µm, in ( B ) 100 µm. The bar graph in ( C ) shows the mean of three independent micrographs ± standard deviation. Asterisks indicate the p -value of the sample compared to the wild-type control. Statistical analysis: One-way ANOVA. ns = non-significant, ** p < 0.01, *** p < 0.001.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Knock-Out, Clone Assay, Staining, Negative Control, Standard Deviation, Control

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: N-glycan structures of C2C12 wild-type and Gne KO myoblasts. MALDI-TOF-TOF MS spectra of N-glycans obtained from panel ( A ) C2C12 wild-type myoblasts and panel ( B ) Gne KO clone #24. Putative cartoon structures assigned were based on composition, tandem mass spectrometry, the literature, and knowledge of N-glycan biosynthetic pathways. All spectra were graphed as % relative intensity. All molecular ions are [M + Na] + . Experiments were repeated on two biological replicates, and the spectra shown are representative. ( C ) MS/MS spectrum of C2C12 wild-type m/z 6431 glycan. ( D ) MS/MS spectrum of C2C12 Gne KO clone #24 m/z 7062 glycan. Isotopic glycan structures with “M” and “m” indicating major (M) and minor (m) abundancies, respectively. Quantification of sialoglycans in two independent experiments indicates that 27.17 ± 2.89% of the detected N-glycans are sialylated in wild-type cells, while there are 12.4 ± 4.05% sialoglycans in Gne KO cells. GlcNAc: N-acetylglucosamine, Man: Mannose, Fuc: Fucose, Gal: Galactose, Neu5Ac: N-acetylneuraminic acid, Neu5Gc: N-glycolylneuraminic acid.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Glycoproteomics, Mass Spectrometry, Tandem Mass Spectroscopy

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: Analysis of glycosylation events in Gne KO clones. ( A ) Endogenous sialic acid ( N -acetylneuraminic acid) biosynthesis pathway. GNE catalyzes the epimerization of UDP-GlcNAc to ManNAc and its subsequent phosphorylation to ManNAc-6-PO 4 . ( B ) Common glycan structures that contain N -acetylglucosamine (GlcNAc). ( C ) Analysis of glycosylation-initiating gene expression in a Sol8 RNAseq data set . ( D ) O-GlcNAcylation in C2C12 myoblasts. ( E ) O-GlcNAcylation in C2C12 myotubes. ( F ) Alcian blue/PAS staining of C2C12 myotubes. ( G ) N-glycan branching via PHA-L staining. GlcNAc: N -acetylglucosamine, GalNAc: N -acetylgalactosamine, PHA-L: phytohaemagglutinin-L, LacNAc: Galβ1-4GlcNAc, LacdiNAc: GalNAcβ1-4GlcNAc. Bar graphs show the mean of three independent experiments ± standard deviation. Asterisks indicate the p -value of the sample compared to the wild-type control. Statistical analysis: One-way ANOVA. ns = non-significant, *** p < 0.001.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Glycoproteomics, Clone Assay, Phospho-proteomics, Gene Expression, Staining, Standard Deviation, Control

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: Expression analysis of muscle-specific genes in C2C12 wild-type and Gne KO cells. qPCR analysis was used to examine gene expression in C2C12 wild-type (WT) and Gne KO (Cl. 24 and Cl. 26) myoblasts and myotubes. Where indicated, differentiation medium was supplemented with 5 mM Neu5Ac for the duration of the entire differentiation protocol. ( A ) mRNA expression of the sodium channel protein type 4 subunit α (Scn4a). ( B ) mRNA expression of voltage-dependent L-type calcium channel subunit alpha-1S (Cacna1s). ( C ) mRNA expression of the ryanodine receptor 1 (Ryr1). ( D ) mRNA expression of the muscle-specific glycogen phosphorylase (Pygm). All values represent the ΔCt value norm. to Gapdh expression. Diff: differentiated, Neu5Ac: N -Acetylneuraminic acid. All graphs represent the mean of three independent experiments ± standard deviation. Asterisks indicate the p -value of the sample compared to the respective condition. Statistical analysis: One-way ANOVA. ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Expressing, Gene Expression, Standard Deviation

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: Reduced glycogen phosphorylase protein expression and decreased ATP production in C2C12 Gne KO cells. ( A ) Western blot analysis of muscle-specific glycogen phosphorylase (Pygm) protein expression in differentiated C2C12 wild-type (WT) and Gne KO clones #24 and #26. ATP-Glo ® assay for the quantitative analysis of intracellular ATP levels in C2C12 wild-type (WT) and Gne KO (Cl. 24 and Cl. 26) ( B ) myoblasts and ( C ) differentiated myotubes, normalized to wild-type cells. RLU: relative luminescent units. Bar graphs show the mean of three independent experiments ± standard deviation. Asterisks indicate the p -value of the sample compared to the respective condition. Statistical analysis: One-way ANOVA, ns = non-significant, *** p < 0.001.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Expressing, Western Blot, Clone Assay, Glo Assay, Standard Deviation

Journal: Cells

Article Title: Gne-Depletion in C2C12 Myoblasts Leads to Alterations in Glycosylation and Myopathogene Expression

doi: 10.3390/cells15020199

Figure Lengend Snippet: Reduced expression of two voltage-dependent L-type calcium channel subunits in C2C12 Gne KO myotubes. ( A ) Western blot showing the protein expression of the voltage-dependent L-type calcium channel subunits α-1s (Cacna1s) and α-2/δ-1 (Cacna2d1). ( B ) Quantitative analysis of Cacna1s protein expression normalized to ponceau and wild-type (WT) expression. ( C ) Quantitative analysis of Cacna2d1 protein expression normalized to ponceau and wild-type (WT) expression. Bar graphs show the mean of three independent experiments ± standard deviation. Asterisks indicate the p -value of the sample compared to the respective condition. Statistical analysis: One-way ANOVA. ns = non-significant, * p < 0.05, *** p < 0.001.

Article Snippet: C2C12 cells (ACC 565) were purchased from DSMZ (Braunschweig, Germany).

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Food science of animal resources

Article Title: Effects on Goat Meat Extracts on α-Glucosidase Inhibitory Activity, Expression of Bcl-2-Associated X (BAX), p53, and p21 in Cell Line and Expression of Atrogin-1, Muscle Atrophy F-Box (MAFbx), Muscle RING-Finger Protein-1 (MuRF-1), and Myosin Heavy Chain-7 (MYH-7) in C2C12 Myoblsts.

doi: 10.5851/kosfa.2023.e6

Figure Lengend Snippet: Fig. 7. Anti-muscular atrophy-related gene levels in C2C12 cells treated with goat hind legs at hydrous extract (HE), hot water extract (HWE), and ethanol extract (EE). (A) Atrogin-1 and (B) myosin heavy chain (MHC) 1b. a,b Values are significantly different (p<0.05).

Article Snippet: Protein extraction and quantification of Atrogin-1, MAFbx, MuRF-1, and MYH-7 expression in C2C12 cells were performed according to the methods described in as above. with the following changes: In order to perform immunoblotting, primary antibodies [rabbit-anti-Atrogin-1 (AP2041, 1:700; ECM Biosciences, Versailles, KY, USA), mouse-anti-MAFbx (sc-166806, 1:300; Santa Cruz Biotechnology), mouse-anti-MuRF-1 (sc-398608, 1:300; Santa Cruz Biotechnology), and mouse-anti-MYH-7 (sc-53089, 1:200; Santa Cruz Biotechnology)] were used.

Techniques:

Journal: Food science of animal resources

Article Title: Effects on Goat Meat Extracts on α-Glucosidase Inhibitory Activity, Expression of Bcl-2-Associated X (BAX), p53, and p21 in Cell Line and Expression of Atrogin-1, Muscle Atrophy F-Box (MAFbx), Muscle RING-Finger Protein-1 (MuRF-1), and Myosin Heavy Chain-7 (MYH-7) in C2C12 Myoblsts.

doi: 10.5851/kosfa.2023.e6

Figure Lengend Snippet: Fig. 8. Anti-muscular atrophy-related gene levels in C2C12 cells treated with goat meat extract. (A) Atrogin-1 and (B) myosin heavy chain (MHC) 1b. a–c Values are significantly different (p<0.05).

Article Snippet: Protein extraction and quantification of Atrogin-1, MAFbx, MuRF-1, and MYH-7 expression in C2C12 cells were performed according to the methods described in as above. with the following changes: In order to perform immunoblotting, primary antibodies [rabbit-anti-Atrogin-1 (AP2041, 1:700; ECM Biosciences, Versailles, KY, USA), mouse-anti-MAFbx (sc-166806, 1:300; Santa Cruz Biotechnology), mouse-anti-MuRF-1 (sc-398608, 1:300; Santa Cruz Biotechnology), and mouse-anti-MYH-7 (sc-53089, 1:200; Santa Cruz Biotechnology)] were used.

Techniques:

Journal: Food science of animal resources

Article Title: Effects on Goat Meat Extracts on α-Glucosidase Inhibitory Activity, Expression of Bcl-2-Associated X (BAX), p53, and p21 in Cell Line and Expression of Atrogin-1, Muscle Atrophy F-Box (MAFbx), Muscle RING-Finger Protein-1 (MuRF-1), and Myosin Heavy Chain-7 (MYH-7) in C2C12 Myoblsts.

doi: 10.5851/kosfa.2023.e6

Figure Lengend Snippet: Fig. 9. Anti-muscular atrophy-related protein levels in C2C12 cells treated with goat meat extract. (A) Atrogin-1, (B) muscle atrophy F- box (MAFbx), (C) muscle RING-finger protein-1 (MuRF-1), and (D) myosin heavy chain-7 (MYH-7). a,b Values are significantly different (p<0.05).

Article Snippet: Protein extraction and quantification of Atrogin-1, MAFbx, MuRF-1, and MYH-7 expression in C2C12 cells were performed according to the methods described in as above. with the following changes: In order to perform immunoblotting, primary antibodies [rabbit-anti-Atrogin-1 (AP2041, 1:700; ECM Biosciences, Versailles, KY, USA), mouse-anti-MAFbx (sc-166806, 1:300; Santa Cruz Biotechnology), mouse-anti-MuRF-1 (sc-398608, 1:300; Santa Cruz Biotechnology), and mouse-anti-MYH-7 (sc-53089, 1:200; Santa Cruz Biotechnology)] were used.

Techniques: