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ATCC cell culture c2c12 murine skeletal muscle myoblasts
Cell Culture C2c12 Murine Skeletal Muscle Myoblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 8324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cell culture c2c12 murine skeletal muscle myoblasts - by Bioz Stars, 2026-06

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(A) Subconfluent C2C12 myoblasts cultured in DMEM were plated in the 24-well base plate at 2.5 × 10 4 /cm 2 . C2C12 myoblasts proliferated for four days and were induced to differentiate in differentiation media (DM), DMEM containing 2% horse serum for 72 hours. On the same day of inducing differentiation, C26 cells and CT26 cells were plated in the co-culture inserts at increasing densities (4.17 × 10 4 /cm 2 , 8.3 × 10 4 /cm 2 , and 1.67 × 10 5 /cm 2 ) and proliferated for 72 hours. C2C12 myoblasts were plated in the inserts at 8.3 × 10 4 /cm 2 as a negative co-culture control. Then, the co-culture inserts containing C26, CT26, or C2C12 cells were moved to the lower compartments of the base plate. Fresh DM was added to both inserts and lower compartments. (B) The co-culture continued for 72 hours, and cells were fixed with 4% paraformaldehyde (PFA) and stained for myosin heavy chain (MyHC, green) and myogenin (MyoG, red). DAPI was used to counterstain the nuclei (blue). Scale bar is set at 200 µm. Myotube diameter (C) , percentage of myotube area (D) , MyoG + /total DAPI + percentage (E) , and total DAPI + cells (F) were quantified to assess potential atrophic effect of C26 cells and CT26 cells. *P<0.05 for pairwise comparison to C2C12 insert control, #P<0.05 for comparison between C26 and CT26 cells.

Journal: bioRxiv

Article Title: C26 and CT26 colorectal cancer models exhibit divergent cachexia phenotypes, intramuscular inflammation, and protein turnover signaling

doi: 10.64898/2026.04.21.719997

Figure Lengend Snippet: (A) Subconfluent C2C12 myoblasts cultured in DMEM were plated in the 24-well base plate at 2.5 × 10 4 /cm 2 . C2C12 myoblasts proliferated for four days and were induced to differentiate in differentiation media (DM), DMEM containing 2% horse serum for 72 hours. On the same day of inducing differentiation, C26 cells and CT26 cells were plated in the co-culture inserts at increasing densities (4.17 × 10 4 /cm 2 , 8.3 × 10 4 /cm 2 , and 1.67 × 10 5 /cm 2 ) and proliferated for 72 hours. C2C12 myoblasts were plated in the inserts at 8.3 × 10 4 /cm 2 as a negative co-culture control. Then, the co-culture inserts containing C26, CT26, or C2C12 cells were moved to the lower compartments of the base plate. Fresh DM was added to both inserts and lower compartments. (B) The co-culture continued for 72 hours, and cells were fixed with 4% paraformaldehyde (PFA) and stained for myosin heavy chain (MyHC, green) and myogenin (MyoG, red). DAPI was used to counterstain the nuclei (blue). Scale bar is set at 200 µm. Myotube diameter (C) , percentage of myotube area (D) , MyoG + /total DAPI + percentage (E) , and total DAPI + cells (F) were quantified to assess potential atrophic effect of C26 cells and CT26 cells. *P<0.05 for pairwise comparison to C2C12 insert control, #P<0.05 for comparison between C26 and CT26 cells.

Article Snippet: The murine skeletal muscle cell line C2C12 (ATCC, CRL-1772) and two colorectal carcinoma cell lines including C26 (a gift from Dr. Keehong Kim at Purdue University) and CT26 (ATCC, CRL-2638) were cultured separately in Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in a cell incubator with 5% CO 2 .

Techniques: Cell Culture, Co-Culture Assay, Control, Staining, Comparison

(A): To assess the effect of H-PUFAs and D-PUFAs during differentiation, confluent C2C12 myoblasts were induced to differentiate for 72 hours in DM supplemented with a 25 µM dose of individual H-PUFAs or D-PUFAs. (B-D): Total DAPI + cell count (B) , average myonuclei per myotube (C) , and average myotube diameter during differentiation (D) were quantified using ImageJ. (E): To assess the effect of H-PUFAs and D-PUFAs on mature myotubes, confluent C2C12 myoblasts were induced to differentiate for 72 hours in DM prior to supplementation with a 25 µM dose of individual H-PUFAs or D-PUFAs for 72 hours. (F-H): Total DAPI + cell count (F) , average myonuclei per myotube (G) , and myotube diameter (H) were quantified using ImageJ. For both experiments, myotubes were fixed by 4% PFA and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. *P<0.05 for difference compared to vehicle, #P<0.05 for difference between H-PUFA and D-PUFA.

Journal: bioRxiv

Article Title: Deuterated Polyunsaturated Fatty Acids Alleviate In Vitro Skeletal Muscle Dysfunction Induced by Oxidative Stress

doi: 10.64898/2025.12.29.696779

Figure Lengend Snippet: (A): To assess the effect of H-PUFAs and D-PUFAs during differentiation, confluent C2C12 myoblasts were induced to differentiate for 72 hours in DM supplemented with a 25 µM dose of individual H-PUFAs or D-PUFAs. (B-D): Total DAPI + cell count (B) , average myonuclei per myotube (C) , and average myotube diameter during differentiation (D) were quantified using ImageJ. (E): To assess the effect of H-PUFAs and D-PUFAs on mature myotubes, confluent C2C12 myoblasts were induced to differentiate for 72 hours in DM prior to supplementation with a 25 µM dose of individual H-PUFAs or D-PUFAs for 72 hours. (F-H): Total DAPI + cell count (F) , average myonuclei per myotube (G) , and myotube diameter (H) were quantified using ImageJ. For both experiments, myotubes were fixed by 4% PFA and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. *P<0.05 for difference compared to vehicle, #P<0.05 for difference between H-PUFA and D-PUFA.

Article Snippet: The C2C12 murine skeletal muscle cell line (ATCC, CRL-1772) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, 11995-065) containing 10% fetal bovine serum (FBS, Corning, 35-010-CV) and antibiotics (penicillin 100 U/mL, streptomycin 100 μg/mL, Gibco, 15140-122) at 37 °C in humid air with 5% CO 2 .

Techniques: Cell Counting, Staining

( A): To assess the effect of H 2 O 2 on mature myotubes, confluent C2C12 myoblasts were induced to differentiate for 48 h in DM and then pre-treated with a 25 µM dose of individual H-PUFA or D-PUFA for a further 24 h. Subsquently, myotubes continued to grow in fresh DM with 2 mM H 2 O 2 and a fresh 25 µM of H-PUFAs or D-PUFAs for 48 hours. Myotubes were fixed in 4% PFA and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. (B-D): Total DAPI + cell count ( B ), average myonuclei per myotube ( C ), and myotube area ( D ) were quantified using ImageJ. ( E): Bodipy C11 staining of C2C12 cells receiving H-PUFA or D-PUFA in the presence of H 2 O 2 . Oxidized lipids are in green and reduced lipids are in red. *P<0.05 for difference of PUFAs compared to H 2 O 2 , #P<0.05 for difference between H-PUFA and D-PUFA.

Journal: bioRxiv

Article Title: Deuterated Polyunsaturated Fatty Acids Alleviate In Vitro Skeletal Muscle Dysfunction Induced by Oxidative Stress

doi: 10.64898/2025.12.29.696779

Figure Lengend Snippet: ( A): To assess the effect of H 2 O 2 on mature myotubes, confluent C2C12 myoblasts were induced to differentiate for 48 h in DM and then pre-treated with a 25 µM dose of individual H-PUFA or D-PUFA for a further 24 h. Subsquently, myotubes continued to grow in fresh DM with 2 mM H 2 O 2 and a fresh 25 µM of H-PUFAs or D-PUFAs for 48 hours. Myotubes were fixed in 4% PFA and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. (B-D): Total DAPI + cell count ( B ), average myonuclei per myotube ( C ), and myotube area ( D ) were quantified using ImageJ. ( E): Bodipy C11 staining of C2C12 cells receiving H-PUFA or D-PUFA in the presence of H 2 O 2 . Oxidized lipids are in green and reduced lipids are in red. *P<0.05 for difference of PUFAs compared to H 2 O 2 , #P<0.05 for difference between H-PUFA and D-PUFA.

Techniques: Staining, Cell Counting

(A) To assess the effect of erastin during differentiation, proliferating C2C12 myoblasts were pretreated with a 25 µM dose of individual D-PUFAs including D-ARA, D-EPA, D-DPA, or D-DHA for 24 hours. Confluent myoblasts were then induced to differentiate in DM containing ferroptosis inducer erastin (10 µM) and fresh D-PUFAs (25 µM) for 72 hours. (B-D): Total DAPI + cell count ( B ), average myonuclei per myotube ( C ), and average myotube diameter ( D ) were measured using ImageJ. (E): To assess the effect of erastin on mature myotubes, confluent C2C12 cells were induced to differentiate in DM for 48 hours and then pretreated with individual D-PUFAs for 24 hours. Subsquently, myotubes continued to grow in fresh DM containing erastin (10 µM) and fresh D-PUFAs for 72 hours. (F-H): Total DAPI + cell count ( F ), average myonuclei per myotube ( G ), and myotube diameter ( H ) were quantified using ImageJ. Myotubes were fixed and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. *P<0.05 for difference during differentiation, #P<0.05 for difference post differentiation.

Journal: bioRxiv

Article Title: Deuterated Polyunsaturated Fatty Acids Alleviate In Vitro Skeletal Muscle Dysfunction Induced by Oxidative Stress

doi: 10.64898/2025.12.29.696779

Figure Lengend Snippet: (A) To assess the effect of erastin during differentiation, proliferating C2C12 myoblasts were pretreated with a 25 µM dose of individual D-PUFAs including D-ARA, D-EPA, D-DPA, or D-DHA for 24 hours. Confluent myoblasts were then induced to differentiate in DM containing ferroptosis inducer erastin (10 µM) and fresh D-PUFAs (25 µM) for 72 hours. (B-D): Total DAPI + cell count ( B ), average myonuclei per myotube ( C ), and average myotube diameter ( D ) were measured using ImageJ. (E): To assess the effect of erastin on mature myotubes, confluent C2C12 cells were induced to differentiate in DM for 48 hours and then pretreated with individual D-PUFAs for 24 hours. Subsquently, myotubes continued to grow in fresh DM containing erastin (10 µM) and fresh D-PUFAs for 72 hours. (F-H): Total DAPI + cell count ( F ), average myonuclei per myotube ( G ), and myotube diameter ( H ) were quantified using ImageJ. Myotubes were fixed and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. *P<0.05 for difference during differentiation, #P<0.05 for difference post differentiation.

Techniques: Cell Counting, Staining

(A): To assess the effect of D-PUFA dose during differentiation, proliferating C2C12 myoblasts were pretreated with an increasing dose of D-ARA, D-ALA, or D-LA (0, 25, 50, 100 µM) for 24 hours. Confluent myoblasts were then induced to differentiate for 72 h in DM containing erastin (10 µM) and fresh D-ARA, D-ALA, or D-LA at respective concentrations. Myotubes were fixed and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. (B-G): Myotube diameter ( B, D, & F ) and average nuclei per myotube ( C, E, & G ) was measured in cultures receiving treatment with D-ARA ( B-C ), D-ALA ( D-E ), and D-LA ( F-G ). (H): Neutral lipid stained by BODIPY 493/503 in the dose response experiment. Representative images shown in (A) and (H ) were taken from the same fields of view. *P<0.05 for difference compared to cells receiving erastin alone.

Journal: bioRxiv

Article Title: Deuterated Polyunsaturated Fatty Acids Alleviate In Vitro Skeletal Muscle Dysfunction Induced by Oxidative Stress

doi: 10.64898/2025.12.29.696779

Figure Lengend Snippet: (A): To assess the effect of D-PUFA dose during differentiation, proliferating C2C12 myoblasts were pretreated with an increasing dose of D-ARA, D-ALA, or D-LA (0, 25, 50, 100 µM) for 24 hours. Confluent myoblasts were then induced to differentiate for 72 h in DM containing erastin (10 µM) and fresh D-ARA, D-ALA, or D-LA at respective concentrations. Myotubes were fixed and visualized after immunofluorescent staining for sarcomeric myosin (MF20c, colored green) and myogenin (F5Dc, colored red). Cell nuclei were counterstained with DAPI (blue). Scale bar is 200 µm. (B-G): Myotube diameter ( B, D, & F ) and average nuclei per myotube ( C, E, & G ) was measured in cultures receiving treatment with D-ARA ( B-C ), D-ALA ( D-E ), and D-LA ( F-G ). (H): Neutral lipid stained by BODIPY 493/503 in the dose response experiment. Representative images shown in (A) and (H ) were taken from the same fields of view. *P<0.05 for difference compared to cells receiving erastin alone.

Techniques: Staining

At 3 days post-differentiation, mature C2C12 myotubes were co-treated with erastin (10 µM) and D-PUFAs (25 µM) in fresh DM for 24 hours. Cell mRNA expression of Scl7a11 ( A ), Hmox1 ( B ), Fbxo32 ( C ), and Il6 ( D ) were assessed using RT-qPCR as described in methods. Gene expression was normalized to Tbp . *P<0.05 for effect of D-PUFAs compared to vehicle, #P<0.05 for effect of erastin.

Journal: bioRxiv

Article Title: Deuterated Polyunsaturated Fatty Acids Alleviate In Vitro Skeletal Muscle Dysfunction Induced by Oxidative Stress

doi: 10.64898/2025.12.29.696779

Figure Lengend Snippet: At 3 days post-differentiation, mature C2C12 myotubes were co-treated with erastin (10 µM) and D-PUFAs (25 µM) in fresh DM for 24 hours. Cell mRNA expression of Scl7a11 ( A ), Hmox1 ( B ), Fbxo32 ( C ), and Il6 ( D ) were assessed using RT-qPCR as described in methods. Gene expression was normalized to Tbp . *P<0.05 for effect of D-PUFAs compared to vehicle, #P<0.05 for effect of erastin.

Techniques: Expressing, Quantitative RT-PCR, Gene Expression

Shows the image of the cell population of the C2C12 cell plates at Magnification: 100×; Scale Bar: 100 μm. KEY : ( A ): C2C12: Normal cells without the myotubes; ( B ): Differentiated cells Positive Control (healthy matured cells) with myotubes; ( C ): Negative control induced with 0.75 mM Palmitate (diabetic cells); ( D ): Induced with 0.75 mM Palmitate + 0.1 µM Metformin (diabetic cells treated with metformin); ( E ): Induced with 0.75 mM Palmitate + PG (diabetic cells treated with 150 µg/mL Solanum kumba ); ( F ): Induced with 0.75 mM Palmitate + PGW (diabetic cells treated with 150 µg/mL Solanum aethiopicum ); ( G ): Healthy matured cells with 150 µg/mL PG ( Solanum kumba ); ( H ): Healthy matured cells with 150 µg/mL PGW ( Solanum aethiopicum ).

Journal: International Journal of Molecular Sciences

Article Title: Eggplant ( Solanum spp.) Fruits Dietary Polyphenols Upregulate the Expression of Glucose Transporter Protein in Palmitate-Induced Diabetic Cell Line C2C12

doi: 10.3390/ijms26167762

Figure Lengend Snippet: Shows the image of the cell population of the C2C12 cell plates at Magnification: 100×; Scale Bar: 100 μm. KEY : ( A ): C2C12: Normal cells without the myotubes; ( B ): Differentiated cells Positive Control (healthy matured cells) with myotubes; ( C ): Negative control induced with 0.75 mM Palmitate (diabetic cells); ( D ): Induced with 0.75 mM Palmitate + 0.1 µM Metformin (diabetic cells treated with metformin); ( E ): Induced with 0.75 mM Palmitate + PG (diabetic cells treated with 150 µg/mL Solanum kumba ); ( F ): Induced with 0.75 mM Palmitate + PGW (diabetic cells treated with 150 µg/mL Solanum aethiopicum ); ( G ): Healthy matured cells with 150 µg/mL PG ( Solanum kumba ); ( H ): Healthy matured cells with 150 µg/mL PGW ( Solanum aethiopicum ).

Article Snippet: Protease tablets were purchased from Roche (South San Francisco, CA, USA), while reverse and forward primers were gotten from Integrated DNA Technologies (Coralville, IA, USA) as well as the reference gene β-actin, Murine C2C12 skeletal muscle cells (CRL 1722) were obtained from the American Type Culture Collection (Manassas, VA, USA) and cell culture kits for cellular antioxidants were supplied by Sigma Aldrich (St. Louis, MO, USA).

Techniques: Positive Control, Negative Control

Intracellular effect of eggplant extracts on the antioxidant enzymes glutathione peroxidase ( A ), catalase ( B ), superoxide dismutase ( C ), and glucose oxidase ( D ) activities in C2C12 cells. Values represent mean ± standard deviation (n = 3). Bars with the same letter are not significant ( p < 0.05). KEY: from the left to the right on the bar chart (Control): is normal cells (Pal): is the diabetic cells without treatment, (Pal + Mef): is diabetic cells with metformin, (Pal + PG): diabetic cells + Solanum kumba , (Pal + PGW): diabetic cells + Solanum aethiopicum , (PG): normal cells + Solanum kumba , (PGW): normal cells + Solanum aethiopicum .

Journal: International Journal of Molecular Sciences

Article Title: Eggplant ( Solanum spp.) Fruits Dietary Polyphenols Upregulate the Expression of Glucose Transporter Protein in Palmitate-Induced Diabetic Cell Line C2C12

doi: 10.3390/ijms26167762

Figure Lengend Snippet: Intracellular effect of eggplant extracts on the antioxidant enzymes glutathione peroxidase ( A ), catalase ( B ), superoxide dismutase ( C ), and glucose oxidase ( D ) activities in C2C12 cells. Values represent mean ± standard deviation (n = 3). Bars with the same letter are not significant ( p < 0.05). KEY: from the left to the right on the bar chart (Control): is normal cells (Pal): is the diabetic cells without treatment, (Pal + Mef): is diabetic cells with metformin, (Pal + PG): diabetic cells + Solanum kumba , (Pal + PGW): diabetic cells + Solanum aethiopicum , (PG): normal cells + Solanum kumba , (PGW): normal cells + Solanum aethiopicum .

Techniques: Standard Deviation, Control

The genetic expression with RT-qPCR on GLUT4 , NRF-1 , and MEF-2A in C2C12 cells.

Journal: International Journal of Molecular Sciences

Article Title: Eggplant ( Solanum spp.) Fruits Dietary Polyphenols Upregulate the Expression of Glucose Transporter Protein in Palmitate-Induced Diabetic Cell Line C2C12

doi: 10.3390/ijms26167762

Figure Lengend Snippet: The genetic expression with RT-qPCR on GLUT4 , NRF-1 , and MEF-2A in C2C12 cells.

Techniques: Expressing, Quantitative RT-PCR

A , D , G DAPI-stained microcarriers showed C2C12 cell adhesion on day 1 and oval nuclei by day 7 in 1G and µG conditions. B , E , H Phalloidin-stained F-actin filaments showed cell body elongation after 7 days, particularly in µG condition, with less pronounced extension in the 1G bioreactors. C , F , I Merged images of DAPI and F-actin staining illustrated overall cell morphology and cytoskeletal structure. J Quantification of nuclei aspect ratio showed increased nuclei elongation in both dynamic conditions, significantly higher in stirred bioreactor. K Quantification of cell body length revealed significant elongation in µG condition. n = 4 and * indicates significant differences at p ≤ 0.05.

Journal: NPJ Science of Food

Article Title: Microcarrier-seeded muscle cells exhibit delayed differentiation in simulated microgravity compared to a terrestrial bioreactor

doi: 10.1038/s41538-025-00498-5

Figure Lengend Snippet: A , D , G DAPI-stained microcarriers showed C2C12 cell adhesion on day 1 and oval nuclei by day 7 in 1G and µG conditions. B , E , H Phalloidin-stained F-actin filaments showed cell body elongation after 7 days, particularly in µG condition, with less pronounced extension in the 1G bioreactors. C , F , I Merged images of DAPI and F-actin staining illustrated overall cell morphology and cytoskeletal structure. J Quantification of nuclei aspect ratio showed increased nuclei elongation in both dynamic conditions, significantly higher in stirred bioreactor. K Quantification of cell body length revealed significant elongation in µG condition. n = 4 and * indicates significant differences at p ≤ 0.05.

Article Snippet: The murine skeletal muscle cell line C2C12 cells (American Type Culture Collection, Manassas, VA, USA) were cultured in regular complete media, comprising Dulbecco’s Modified Eagle Medium (DMEM) (Gibco) with 10% fetal bovine serum (FBS) (Gibco), 1% penicillin/streptomycin (Gibco), and 1% GlutaMAX (Gibco), at 37 °C in a humidified 5% CO 2 incubator until they reached 70–80% confluency.

Techniques: Staining

A , C Representative flow cytometry images of C2C12 cells at Day 0 and Day 8 in 1G and µG bioreactors. B , D Quantitative analysis of the percentage of MYHC- and MYOD-positive cells at Day 0 and Day 8 revealed a higher decrease in MYOD-positive cells and a lower increase in MYHC-positive cells in the µG bioreactor compared to the 1G bioreactor. This suggests a delayed transition from the proliferative phase to differentiation in µG, with impaired muscle cell differentiation. n = 3 and * indicates significant differences at p ≤ 0.05.

Journal: NPJ Science of Food

Article Title: Microcarrier-seeded muscle cells exhibit delayed differentiation in simulated microgravity compared to a terrestrial bioreactor

doi: 10.1038/s41538-025-00498-5

Figure Lengend Snippet: A , C Representative flow cytometry images of C2C12 cells at Day 0 and Day 8 in 1G and µG bioreactors. B , D Quantitative analysis of the percentage of MYHC- and MYOD-positive cells at Day 0 and Day 8 revealed a higher decrease in MYOD-positive cells and a lower increase in MYHC-positive cells in the µG bioreactor compared to the 1G bioreactor. This suggests a delayed transition from the proliferative phase to differentiation in µG, with impaired muscle cell differentiation. n = 3 and * indicates significant differences at p ≤ 0.05.

Techniques: Flow Cytometry, Cell Differentiation

SCE changes cell morphology during myogenic differentiation. C2C12 myoblasts were cultured in growth medium until 80% confluence and then changed to DMEM containing 2% horse serum and 1, 5 and 10 ng/ml SCE for 5 days. The medium was changed every 2 days and fresh SCE was added. (A) Cell viability was assessed using the XTT assay (n=6 per group). (B) Images of morphological changes were obtained using light microscopy at the end of the experiment (5 days) (n=4 per group). Bar, 50 µm. SCE, Saururus chinensis (Lour.) Baill. Extract.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE changes cell morphology during myogenic differentiation. C2C12 myoblasts were cultured in growth medium until 80% confluence and then changed to DMEM containing 2% horse serum and 1, 5 and 10 ng/ml SCE for 5 days. The medium was changed every 2 days and fresh SCE was added. (A) Cell viability was assessed using the XTT assay (n=6 per group). (B) Images of morphological changes were obtained using light microscopy at the end of the experiment (5 days) (n=4 per group). Bar, 50 µm. SCE, Saururus chinensis (Lour.) Baill. Extract.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Cell Culture, XTT Assay, Light Microscopy

SCE promotes myotube formation by C2C12 cells. C2C12 myoblasts were cultured in a DMEM containing 2% horse serum and SCE (0,1, 5 and 10 ng/ml) for 5 days. (A) Immunofluorescence staining for the myotube marker, MyHC (green), and the nuclei marker, DAPI (blue), revealed myotube formation by C2C12 cells (n=4 per group). (B) Myotube diameters in randomly selected fields were quantified using an image analysis program. ***P<0.01 vs. the control group. Scale bar, 50 µm. SCE, Saururus chinensis (Lour.) Baill. Extract.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE promotes myotube formation by C2C12 cells. C2C12 myoblasts were cultured in a DMEM containing 2% horse serum and SCE (0,1, 5 and 10 ng/ml) for 5 days. (A) Immunofluorescence staining for the myotube marker, MyHC (green), and the nuclei marker, DAPI (blue), revealed myotube formation by C2C12 cells (n=4 per group). (B) Myotube diameters in randomly selected fields were quantified using an image analysis program. ***P<0.01 vs. the control group. Scale bar, 50 µm. SCE, Saururus chinensis (Lour.) Baill. Extract.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Cell Culture, Immunofluorescence, Staining, Marker, Control

SCE increases the expression of muscle-specific factors related to myogenesis. C2C12 myoblasts were induced to differentiate in DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5 and 10 ng/ml), and analyzed by (A) RT-qPCR (n=3 per group) and (B) western blotting (n=3 per group). The expression of MyHC, MyoD, myogenin and Myf5 over the time course of myogenesis was detected through (C) RT-qPCR (n=3 per group) and (D) western blotting (n=3 per group) in cells treated with SCE. *P<0.05, **P<0.01 and ***P<0.001 vs. the control group; #P<0.05, ##P<0.01 and ###P<0.001 vs. the SCE-treated group at the corresponding indicated times. Non-β-catenin and β-catenin were detected by western blotting depending on (E) the concentration or (F) the time of SCE. SCE, Saururus chinensis (Lour.) Baill. extract; RT-qPCR, reverse transcription-quantitative PCR; MyHC, myosin heavy chain; MyoD, myogenic differentiation 1; Myf5, Myogenic Factor 5.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE increases the expression of muscle-specific factors related to myogenesis. C2C12 myoblasts were induced to differentiate in DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5 and 10 ng/ml), and analyzed by (A) RT-qPCR (n=3 per group) and (B) western blotting (n=3 per group). The expression of MyHC, MyoD, myogenin and Myf5 over the time course of myogenesis was detected through (C) RT-qPCR (n=3 per group) and (D) western blotting (n=3 per group) in cells treated with SCE. *P<0.05, **P<0.01 and ***P<0.001 vs. the control group; #P<0.05, ##P<0.01 and ###P<0.001 vs. the SCE-treated group at the corresponding indicated times. Non-β-catenin and β-catenin were detected by western blotting depending on (E) the concentration or (F) the time of SCE. SCE, Saururus chinensis (Lour.) Baill. extract; RT-qPCR, reverse transcription-quantitative PCR; MyHC, myosin heavy chain; MyoD, myogenic differentiation 1; Myf5, Myogenic Factor 5.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Concentration Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

SCE upregulates the expression of biogenesis-regulating factors during C2C12 differentiation. C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5 and 10 ng/ml) and analyzed using (A and E) RT-qPCR (n=3 per group) and (B) western blotting (n=3 per group). Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and subjected to (C and F) RT-qPCR (n=3 per group) and (D) western blotting (n=3 per group). GAPDH was used as a loading control. **P<0.01 and ***P<0.001 vs. the control group; ##P<0.01 and ###P<0.001 vs. the SCE-treated group at the corresponding indicated times. SCE, Saururus chinensis (Lour.) Baill. extract; RT-qPCR, reverse transcription-quantitative PCR; PGC-1a, peroxisome proliferator-activated receptor-gamma coactivator-1 α; NRF1, nuclear respiratory factor 1.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE upregulates the expression of biogenesis-regulating factors during C2C12 differentiation. C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5 and 10 ng/ml) and analyzed using (A and E) RT-qPCR (n=3 per group) and (B) western blotting (n=3 per group). Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and subjected to (C and F) RT-qPCR (n=3 per group) and (D) western blotting (n=3 per group). GAPDH was used as a loading control. **P<0.01 and ***P<0.001 vs. the control group; ##P<0.01 and ###P<0.001 vs. the SCE-treated group at the corresponding indicated times. SCE, Saururus chinensis (Lour.) Baill. extract; RT-qPCR, reverse transcription-quantitative PCR; PGC-1a, peroxisome proliferator-activated receptor-gamma coactivator-1 α; NRF1, nuclear respiratory factor 1.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

SCE activates the AMPK-HDAC5 pathways in myotubes. (A and C) C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5, 10 ng/ml) and analyzed by western blotting (upper panel). Quantification of protein expression levels (n=3 per group) (lower panel). *P<0.05 and ***P<0.01 vs. the control. (B and D) Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and western blotting was performed using the indicated antibodies (upper panel). Quantification of protein expression levels (n=3 per group) (down). *P<0.05 and ***P<0.01 vs. the DMSO; ###P<0.001 vs. the DMSO at the indicated time points. SCE, Saururus chinensis (Lour.) Baill. extract; AMPK, AMP-activated protein kinase; HDAC5, histone deacetylase 5; p-, phosphorylated.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE activates the AMPK-HDAC5 pathways in myotubes. (A and C) C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentrations of SCE (1, 5, 10 ng/ml) and analyzed by western blotting (upper panel). Quantification of protein expression levels (n=3 per group) (lower panel). *P<0.05 and ***P<0.01 vs. the control. (B and D) Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and western blotting was performed using the indicated antibodies (upper panel). Quantification of protein expression levels (n=3 per group) (down). *P<0.05 and ***P<0.01 vs. the DMSO; ###P<0.001 vs. the DMSO at the indicated time points. SCE, Saururus chinensis (Lour.) Baill. extract; AMPK, AMP-activated protein kinase; HDAC5, histone deacetylase 5; p-, phosphorylated.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Western Blot, Expressing, Control, Histone Deacetylase Assay

SCE regulates AKT/mTOR and its downstream effectors in myotubes. (A) C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentration of SCE (1, 5 and 10 ng/ml) and analyzed using western blotting (left). Quantification of protein expression levels (n=3 per group) (right). **P<0.01 and ***P<0.001 vs. the control. (B) Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and western blotting was performed using the indicated antibodies (left). Quantification of protein expression levels (n=3 per group) (right). *P<0.05, **P<0.01 and ***P<0.001 vs. the DMSO; #P<0.05, ##P<0.01 and ###P<0.001 vs. the DMSO at the indicated time points. SCE, Saururus chinensis (Lour.) Baill. extract; p-, phosphorylated; p70S6K1, ribosomal protein S6 kinase B1.

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Figure Lengend Snippet: SCE regulates AKT/mTOR and its downstream effectors in myotubes. (A) C2C12 myoblasts were induced to differentiate in a DMEM containing 2% horse serum, treated for 5 days with different concentration of SCE (1, 5 and 10 ng/ml) and analyzed using western blotting (left). Quantification of protein expression levels (n=3 per group) (right). **P<0.01 and ***P<0.001 vs. the control. (B) Cells were treated with 10 ng/ml SCE in DM for 1, 3 and 5 days and western blotting was performed using the indicated antibodies (left). Quantification of protein expression levels (n=3 per group) (right). *P<0.05, **P<0.01 and ***P<0.001 vs. the DMSO; #P<0.05, ##P<0.01 and ###P<0.001 vs. the DMSO at the indicated time points. SCE, Saururus chinensis (Lour.) Baill. extract; p-, phosphorylated; p70S6K1, ribosomal protein S6 kinase B1.

Article Snippet: Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Concentration Assay, Western Blot, Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Cell Culture, XTT Assay, Light Microscopy

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Cell Culture, Immunofluorescence, Staining, Marker, Control

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Concentration Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Western Blot, Expressing, Control, Histone Deacetylase Assay

Journal: Molecular Medicine Reports

Article Title: Saururus chinensis (Lour.) Baill. extract promotes skeletal muscle cell differentiation by positively regulating mitochondrial biogenesis and AKT/mTOR signaling in vitro

doi: 10.3892/mmr.2024.13250

Article Snippet: Cell culture Murine C2C12 skeletal muscle cell line (cat. no. CRL-1772) were purchased from American Type Culture Collection.

Techniques: Concentration Assay, Western Blot, Expressing, Control

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