c2c12 mouse myoblast cell line  (Procell Inc)

Journal: Toxicology Reports

Article Title: Chlorpyrifos abolished C2C12 myoblast cell proliferation and differentiation via mitochondrial stress

doi: 10.1016/j.toxrep.2025.102041

Figure Lengend Snippet: Cytotoxicity of Chlorpyrifos (CPF) on C2C12 myoblast cells. Subconfluent C2C12 myoblasts were cultured in low-serum medium containing CPF at 0–100 µM for 24–72 h. Cell viability was determined by MTT assay (A) and cell morphology was photographed (B). ** P < 0.01, *** P < 0.001 compared with control. Scale bar = 200 µm.

Article Snippet: The C2C12 mouse myoblast cell line was purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA), and was maintained in growth medium (GM) composed of DMEM supplemented with 10 % foetal bovine serum and 1 % antibiotics at 37 °C in a humidified CO 2 incubator set to 5 % CO 2 .

Techniques: Cell Culture, MTT Assay, Control

Journal: Toxicology Reports

Article Title: Chlorpyrifos abolished C2C12 myoblast cell proliferation and differentiation via mitochondrial stress

doi: 10.1016/j.toxrep.2025.102041

Figure Lengend Snippet: Chlorpyrifos (CPF) stimulated reactive oxygen species (ROS) production. C2C12 myoblasts were cultured in CPF at indicated concentrations for 72 h. ROS production was observed by H 2 DCFDA assay (green, arrowhead) (A) and fluorescence intensity was measured (B). Expression of mitochondrial-related genes was determined by real-time PCR (C). Expression of apoptotic markers was determined by western blotting (D) and band intensity was measured (E). * P < 0.05, ** P < 0.01, *** P < 0.001 compared with control. Scale bar = 50 µm.

Article Snippet: The C2C12 mouse myoblast cell line was purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA), and was maintained in growth medium (GM) composed of DMEM supplemented with 10 % foetal bovine serum and 1 % antibiotics at 37 °C in a humidified CO 2 incubator set to 5 % CO 2 .

Techniques: Cell Culture, Fluorescence, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Toxicology Reports

Article Title: Chlorpyrifos abolished C2C12 myoblast cell proliferation and differentiation via mitochondrial stress

doi: 10.1016/j.toxrep.2025.102041

Figure Lengend Snippet: Chlorpyrifos (CPF) inhibited C2C12 myoblast proliferation. Subconfluent C2C12 myoblasts were cultured in growth medium containing CPF at 0–100 µM for 24 and 48 h. Cell proliferation was determined by MTT assay (A) and cell morphology at 48 h after treatment was photographed (B). * P < 0.05, ** P < 0.01, *** P < 0.001 compared with control. Scale bar = 200 µm.

Article Snippet: The C2C12 mouse myoblast cell line was purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA), and was maintained in growth medium (GM) composed of DMEM supplemented with 10 % foetal bovine serum and 1 % antibiotics at 37 °C in a humidified CO 2 incubator set to 5 % CO 2 .

Techniques: Cell Culture, MTT Assay, Control

Journal: Toxicology Reports

Article Title: Chlorpyrifos abolished C2C12 myoblast cell proliferation and differentiation via mitochondrial stress

doi: 10.1016/j.toxrep.2025.102041

Figure Lengend Snippet: Chlorpyrifos (CPF) stimulated myoblast cell cycle arrest. Subconfluent C2C12 myoblasts were cultured in growth medium containing CPF at indicated concentrations for 48 h. Cell proliferation was determined by cell counting (A) and Ki-67 immunostaining (red, arrowhead) (B). Cell cycle distribution was determined by flow cytometry (C) and percentage of cells at each stage was calculated (D). Protein expression was determined by western blotting (E) and band intensity was measured (F). * P < 0.05, ** P < 0.01, *** P < 0.001 compared with control. Scale bar = 50 µm.

Article Snippet: The C2C12 mouse myoblast cell line was purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA), and was maintained in growth medium (GM) composed of DMEM supplemented with 10 % foetal bovine serum and 1 % antibiotics at 37 °C in a humidified CO 2 incubator set to 5 % CO 2 .

Techniques: Cell Culture, Cell Counting, Immunostaining, Flow Cytometry, Expressing, Western Blot, Control

Journal: Toxicology Reports

Article Title: Chlorpyrifos abolished C2C12 myoblast cell proliferation and differentiation via mitochondrial stress

doi: 10.1016/j.toxrep.2025.102041

Figure Lengend Snippet: Chlorpyrifos (CPF) abrogated C2C12 myoblast differentiation. Confluent C2C12 myoblasts were switched into differentiation medium containing CPF at 0–25 µM for 72 h. Myotubes were photographed and stained with anti-myosin heavy chain antibody (green, arrowhead) (A). Myotube size was measured (B). Protein expression was evaluated by western blotting (C, E) and band intensity was measured (D, F). * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar = 50 µm.

Article Snippet: The C2C12 mouse myoblast cell line was purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA), and was maintained in growth medium (GM) composed of DMEM supplemented with 10 % foetal bovine serum and 1 % antibiotics at 37 °C in a humidified CO 2 incubator set to 5 % CO 2 .

Techniques: Staining, Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Glucose, glutamine, lactic acid and α-ketoglutarate restore metabolic disturbances and atrophic changes in energy-deprived muscle cells

doi: 10.3892/mmr.2025.13562

Figure Lengend Snippet: Effect of single nutrient supplementation on metabolic activity. (A) Cells were incubated in regular DMEM supplemented with the indicated nutrient for 24 h. (B and C) C2C12 myotubes were cultured in starvation medium only or with the indicated nutrient for (B) 5 or (C) 24 h. Albumin and NaOH were added as vehicle controls in both regular and starvation media in the experiment for the fatty acid (PA and OA). Metabolic activity was assessed using the MTT assay. Values are expressed as fold-change compared with the values of the control cells incubated in regular DMEM. Values are presented as the mean ± SEM (n=3-4). ***P<0.001 vs. vehicle control; § P<0.05, §§ P<0.01, §§§ P<0.001 vs. starved cells in the same group. Cont, control; Vehicle, vehicle control; Stv, starvation; Glc, glucose; Gln, glutamine; Glu, glutamic acid; Leu, leucine; Val, valine; LA, lactate; βOHB, β-hydroxy butyric acid; αKG, α-ketoglutarate; PA, palmitic acid; OA, oleic acid.

Article Snippet: Mouse C2C12 myoblast cell line (RCB0987) was purchased from the RIKEN BioResource Research Center (Tsukuba, Japan).

Techniques: Activity Assay, Incubation, Cell Culture, MTT Assay, Control

Journal: Molecular Medicine Reports

Article Title: Glucose, glutamine, lactic acid and α-ketoglutarate restore metabolic disturbances and atrophic changes in energy-deprived muscle cells

doi: 10.3892/mmr.2025.13562

Figure Lengend Snippet: Effect of single nutrient supplementation on ATP production. C2C12 myotubes were incubated in starvation medium with or without the indicated nutrients for 24 h. The ATP production rates from glycolysis (Glyco-ATP) and OxPhos (OxPhos-ATP) were determined. Values are expressed as fold-change compared with the values of the control cells incubated in regular DMEM. Values are presented as mean ± SEM (n=3-4). **P<0.01, ***P<0.001 vs. control cells for total ATP production; §§§ P<0.001 vs. starved cells for Glyco-ATP production. Con, control; Glyco-ATP, glycolytic ATP; OxPhos, oxidative phosphorylation; Stv, starvation.

Article Snippet: Mouse C2C12 myoblast cell line (RCB0987) was purchased from the RIKEN BioResource Research Center (Tsukuba, Japan).

Techniques: Incubation, Control, Phospho-proteomics

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression of DDIT4, mTOR, GLUT4, PI3K, and AKT in MHY1485-treated IR C2C12 cells. (A) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, AKT after the activation of mTOR. (B) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, AKT and protein expression levels of DDIT4, GLUT4 after MHY1485 treatment. (C) Glucose levels in culture medium after MHY1485 treatment. (D) Cell TG levels after MHY1485 treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: The C2C12 mouse myoblast cell line (cat. no. CL-0044; Procell Life Science & Technology Co., Ltd.) was cultured in culture medium (cat. no. CM-0044 Procell Life Science & Technology Co., Ltd.) until the cells reached 70-80% confluence.

Techniques: Expressing, Activation Assay, Control

Journal: Biomedical Reports

Article Title: Resveratrol ameliorates high‑fat diet‑induced insulin resistance via the DDIT4/mTOR pathway in skeletal muscle

doi: 10.3892/br.2025.1977

Figure Lengend Snippet: Expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT in DDIT4-siRNA-treated IR C2C12 cells. (A) DDIT4-siRNA-1 had the most significant silencing effect. (B) Protein expression levels of DDIT4, mTOR, GLUT4, PI3K, and AKT after DDIT4-siRNA treatment. (C) Semi-quantification of p-protein/t-protein ratios for mTOR, PI3K, and AKT and protein expression levels of DDIT4 and GLUT4 after DDIT4-siRNA treatment. (D) Glucose levels in culture medium after DDIT4-siRNA treatment. (E) TG levels of C2C12 cells after DDIT4-siRNA treatment. Data are presented as the mean ± SD (n=3). * P<0.05 vs. the CON group; # P<0.05 vs. the PA group; & P<0.05 vs. the PA + RSV group. DDIT4, DNA-damage-inducible transcript 4; mTOR, mammalian target of rapamycin; GLUT4, glucose transporter 4; PI3K, phosphatidylinositol 3-kinase; AKT, protein kinase B; siRNA, small interfering RNA; IR, insulin resistance; p-, phosphorylated; t-, total; TG, triglyceride; CON, control; PA, palmitic acid; RSV, resveratrol.

Article Snippet: The C2C12 mouse myoblast cell line (cat. no. CL-0044; Procell Life Science & Technology Co., Ltd.) was cultured in culture medium (cat. no. CM-0044 Procell Life Science & Technology Co., Ltd.) until the cells reached 70-80% confluence.

Techniques: Expressing, Small Interfering RNA, Control