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: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: Cap1 mRNA and protein levels are downregulated during myogenic differentiation. (A,E) Bright-field images (×20) of murine C2C12 (A) and human LHCN-M2 (E) cells upon differentiation for 4 and 6 days (d4 and d6), in comparison to undifferentiated control cells (d0). Cells were stained with crystal violet. (B,F) Relative Cap1 mRNA in differentiating C2C12 (B) and LHCN-M2 (F) cells, quantified by qRT-PCR and normalized to a set of housekeeping mRNAs. (C,G) Immunoblots of lysates from differentiating C2C12 (C) and LHCN-M2 (G) cells, using antibodies against myosin heavy chain polypeptides 1, 2, 4, and 6 (MYH), CAP1 and GAPDH as a control. (D,H) Quantification of CAP1 immunoblots at myogenic differentiation for 4 and 6 days, normalized to undifferentiated control cells. Error bars , SEM ( n = 3); ** p < 0.01, *** p < 0.001 (Student’s t -test). Scale bar, 200 μm.

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: Comparison, Control, Staining, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: Knockout of Cap1 results in increased size of cells and nuclei and accumulated F-actin fibers in C2C12 cells. (A) Validation of partial knockout by CRISPR-Cas9 (sg-Cap1) and overexpression (dsRed-Cap1) in C2C12 pools by immunoblot, compared to control cells infected with Cas9 only (Cas9). The sg-Cap1 cells show reduced expression of endogenous CAP1 while ectopically expressed dsRed-Cap1 results in an additional band corresponding to tagged CAP1 proteins ( n = 3). (B) Quantification of the cell area covered, based on crystal violet staining ( n = 50 cells). (C) Quantification of the nucleus size ( n = 100 nucleus). (D) Micrographs, the upper panel shows fluorescence images of phalloidin-stained cells. The lower panel shows bright-field images (×20) of crystal violet stained cells. Data presented here are from two-week post transduction. The cell and nucleus size quantifications are presented as box plot, showing mean (cross), median (line), 25th and 75th percentile (box). The whiskers extend to the most extreme data points not considered outliers, and the outliers are represented as dots; *** p < 0.001 (Student’s t -test). Scale bar, 200 μm.

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: Knock-Out, Biomarker Discovery, CRISPR, Over Expression, Western Blot, Control, Infection, Expressing, Staining, Fluorescence, Transduction

Journal: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: Timely downregulation of CAP1 is important for myoblast fusion. (A) Cas9 control cells, knockout (sg-Cap1) and overexpressing (dsRed-Cap1) cells at day 0 (d0), day 4 (d4) and day 6 (d6) of their differentiation. Pools of C2C12 cells were stained for MYH (green) and nucleus (DAPI, orange). (B) Quantification of the percentage of myotubes containing the indicated number of nuclei per myotube in control, knockout and overexpressing myotubes after 6 days of differentiation (minimum 200 MYH positive myotubes were counted). (C) Western blot for the myogenic marker MYH, TUBULIN, MYOG, MYOD, Flag, CAP1, and GAPDH at day 0 (d0), days 4 (d4) and 6 (d6) of the differentiation. (D–G) Quantification of MYH, MYOG, MYOD, and CAP1 immunoblots at myogenic differentiation for 4 and 6 days, normalized to Cas9 control cells. Error bars , SEM ( n = 3); * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t -test). Scale bar, 200 μm.

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: Control, Knock-Out, Staining, Western Blot, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: miRNA (miR-1, miR-133, and miR-206) regulate the expression of Cap1 in murine and human myoblast. (A) The abundance of the indicated miRNAs in total lysates of undifferentiated and differentiated C2C12, determined by RNA-Seq. CPM; counts per million ( n = 3). (B) Schematic of the 3′-UTR of murine Cap1 with the STOP-codon at position 1 and the polyadenylation signal at 1,020 and 1,058 bp. Predicted binding sites for miR-1, miR-133 and miR-206 are indicated by yellow boxes. (C,D) Cap1 mRNA expression in undifferentiated C2C12 (C) and LHCN-M2 (D) cells transfected with the indicated miRNA mimic for 72 h ( n = 3). (E,F) Representative immunoblots of cells transfected with the indicated miRNA. (G,H) Quantification of the CAP1 protein from three independent experiments. Error bars, SEM ( n = 3); * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t -test).

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: Expressing, RNA Sequencing, Binding Assay, Transfection, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: Requirement of the 3′-UTR for Cap1 regulation during myogenesis. (A) The 3′-UTR of Cap1 was deleted in C2C12 cells using CRISPR/Cas9. Δ3′-UTR (Δ) and Cas9-only (C) control cells were differentiated for the indicated times. Representative immunoblots for MYH and CAP1 are shown. (B) Quantification of CAP1 protein at day 0, day 4 and day 6 of differentiation normalized to day 0. (C) Quantification of MYH, normalized to Cas9 control cells at day 4. (D) Immunofluorescence staining (×20) of C2C12 cells stained with MYH (green) and DAPI (orange) after differentiation for 4 and 6 days (d4 and d6), in comparison to undifferentiated control cells (d0). Thick and multinucleated myotubes are reduced in the Δ3′-UTR cells (right panel) at day 6 of differentiation, compared to the Cas9 control (left panel). Error bars, SEM ( n = 3); * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t -test). Scale bar, 200 μm.

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: CRISPR, Control, Western Blot, Immunofluorescence, Staining, Comparison

Journal: Frontiers in Cell and Developmental Biology

Article Title: miRNA mediated downregulation of cyclase-associated protein 1 (CAP1) is required for myoblast fusion

doi: 10.3389/fcell.2022.899917

Figure Lengend Snippet: Model depicting the regulatory circuitry of myogenic C2C12 differentiation via post-transcriptional Cap1 regulation. Under physiological conditions, a timely and necessary downregulation of the Cap1 during myogenesis, is induced by myogenic miRNAs miR-1a-3p, miR-133a-3p, and miR-206-3p whose expression increases manifold during differentiation. The decreased levels of CAP1 increases the F-actin levels that enable myoblasts for elongation, migration and fusion necessary for the myoblasts fusion and myotube maturation. Under experimental conditions, both at the induced overexpression and knockout scenario (on the right side) a decreased fusion index was observed as measured by the thickness of the myotubes as well as the number of the nuclei present in the myosin heavy chain positive myotubes. Endogenous deletion of the Cap1 3′ UTR (on the left side) also resulted in the diminished fusion index similar to the CAP1 overexpressing myoblast. Overall, a timely decrease in the expression of the CAP1 is necessary for myoblast fusion.

Article Snippet: C2C12 (DSMZ—German Collection of Microorganisms and Cell Cultures) was cultured subconfluently in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM sodium pyruvate, and antibiotic-antimycotic (Thermo Fisher) at 37°C and 5% CO 2 .

Techniques: Expressing, Migration, Over Expression, Knock-Out

Journal: Journal of cell science

Article Title: Patch repair protects cells from the small pore-forming toxin aerolysin.

doi: 10.1242/jcs.261018

Figure Lengend Snippet: Fig. 6. Annexins minimally resist aerolysin with limited shedding. HeLa cells were transfected with control (CTL), A1, A2 or A6 siRNA for 3 days and then (A,B) lysed for western blot analysis, or (C) challenged with 31–2000 HU/ml aerolysin (Aero), SLO or ILY for 30 min at 37°C. PI uptake was analyzed by flow cytometry. (A) Portions of the blot were probed with antibodies against A1, A2, A6 or β-actin followed by HRP-conjugated secondary antibodies. (B) Quantification of the knockdown efficiency compared to control siRNA is shown. (D,E) HeLa cells were transfected with A6–YFP and challenged with sublytic toxin concentrations (62 HU/ml pro-aerolysin or aerolysin, or 250 HU/ml SLO) in imaging buffer. The cells were imaged by confocal microscopy for ∼45 min at 37°C and then lysed with 1% Triton X-100. The time to half maximal (t1/2) (D) cytosolic depletion or (E) membrane accumulation of A6 is shown. (F) Shedding of annexin A6+ microvesicles was manually counted and expressed as microvesicles shed/cell/min. (G) HeLa cells were challenged for 15 min at 37°C with sublytic doses of aerolysin, SLO, mass equivalent of AeroY221G or sufficient trypsin to activate pro-aerolysin. Cells were pelleted at 2000 g for 5 min to yield cell pellets (denoted C). Cell supernatants were spun at 100,000 g for 40 min at 4°C and the microvesicle pellet (MV) collected. Fractions were analyzed by western blotting using anti-aerolysin, 6D11 anti- SLO, CPTC-A1-3 anti-annexin A1, anti-Alix, MANLAC-4A7 anti-lamin A/C or AC-15 anti-β-actin. Graphs show the mean (C,F) or geometric mean (D,E). Data points represent (C) individual experiments or (D,E) individual cells from three independent experiments. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, not significant (one-way ANOVA with Tukey post test). Asterisks in G indicates non-specific bands.

Article Snippet: C2C12 myoblasts stably expressing control (ATCC CRL-3419) or Dysf shRNA (ATCC CRL-3418) were cultured in DMEM supplemented with 10% Equafetal serum blend and 2 μg/ml puromycin.

Techniques: Transfection, Control, Western Blot, Flow Cytometry, Knockdown, Imaging, Confocal Microscopy, Membrane

Journal: Journal of cell science

Article Title: Patch repair protects cells from the small pore-forming toxin aerolysin.

doi: 10.1242/jcs.261018

Figure Lengend Snippet: Fig. 7. Patch repair protects cells from aerolysin. (A) HeLa, C2C12 control and Dysf shRNA cells were challenged with 15–1000 HU/ml aerolysin (Aero) or 31–2000 HU/ml SLO for 30 min at 37°C. PI uptake was analyzed by flow cytometry. HeLa cells were either untransfected or transfected with GFP or GFP–Dysferlin (GFP–Dysf) for 48 h and challenged with aerolysin (Aero), SLO or ILY at 31-2000 HU/ml for 30 min at 37°C either in (B,C) 2 mM CaCl2- or (C) 2 mM EGTA- supplemented RPMI. PI uptake was analyzed by flow cytometry. The LC50 was calculated as described in the Materials and Methods. (D,E) GFP–Dysf (green) transfected HeLa cells were challenged with sublytic toxin doses (250 HU/ml SLO or ILY, 62 HU/ml aerolysin or mass equivalent to aerolysin of mutant AeroY221G) in imaging buffer either supplemented with (D) 2 mM CaCl2 or (E) 2 mM EGTA, imaged for ∼45 min at 37°C, and then lysed with 1% Triton X-100. Depletion of GFP–Dysf-containing vesicles from cytosol are shown by arrowheads. Graphs show the mean±s.e.m. of (A) four, (B) seven or (C) five independent experiments. The dotted line indicates the limit of detection. Points on this line had LC50>2000 HU/ml. Micrographs show representative images from three independent experiments. **P<0.01, ***P<0.001, ****P<0.0001 (one- way ANOVA with Tukey post test). Scale bars: 5 μm.

Article Snippet: C2C12 myoblasts stably expressing control (ATCC CRL-3419) or Dysf shRNA (ATCC CRL-3418) were cultured in DMEM supplemented with 10% Equafetal serum blend and 2 μg/ml puromycin.

Techniques: Control, shRNA, Flow Cytometry, Transfection, Mutagenesis, Imaging

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:

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