mouse skeletal muscle c2c12 cells  (ATCC)

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Fibre Type–Specific Proteomics Reveals Shared and Distinct Skeletal Muscle Adaptations to Resistance Training and Beta 2 ‐Adrenergic Agonist

doi: 10.1002/jcsm.70175

Figure Lengend Snippet: Mustn1 and S100a13 are important for cell growth. (a) Log 2 fold changes for MUSTN1 and S100A13 in human muscle samples. (b) Overview of cell experiment. C2C12 cells were treated with either small interfering RNA (siRNA) for Mustn1 or S100a13 at Day 2 of differentiation. (c) Representative pictures of myofibers at Day 6 of differentiation. (d) mRNA expression of Mustn1. (e) mRNA expression of S100a13. (f) Fusion index (number of nuclei in myotubes divided by total number of nuclei). Stars denote p value for comparisons shown by brackets (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: C2C12 muscle cells were obtained from American Type Culture Collection (ATCC CRL‐1772) and grown in Dulbecco's modified Eagle's medium (DMEM), supplemented with 100 U/mL penicillin, 100 μg/mL of streptomycin and 10% fetal bovine serum.

Techniques: Small Interfering RNA, Expressing

Journal: RSC Chemical Biology

Article Title: Signatures of native-like glycosylation in RNA replicon-derived HIV-1 immunogens

doi: 10.1039/d5cb00165j

Figure Lengend Snippet: Differential glycan composition on Env expressed in different cell lines. Glycan composition at reporter glycosylation sites predominantly displaying: oligomannose-type glycans (N332), a mixture of complex- and oligomannose-type glycans (N355), and complex-type glycans (N88). Four different production systems were used as described in the key, including expression in three different cell lines, HEK293F, C2C12 and DC2.4 cells. All the glycan composition observed in the site-specific analysis are shown on the left, simplified in distinct categories represented as, core, oligomannose-type, and complex-type glycans. Some compositions annotated as complex-type glycans can exhibit isomers formally constituting hybrid-type glycans. Represented glycan compositions (See Methods for full classification) are colored according to the scale provided in the right for each category of glycan composition. The representative glycan composition data at all the sites is the average of three or more biological replicates.

Article Snippet: The mouse muscle cell line (C2C12) was cultured in Dulbecco's modified Eagles medium supplemented with 10% fetal bovine serum (FBS) as recommended in manufacturer's protocol (American type culture collection, Catalogue no. CRL-1772).

Techniques: Glycoproteomics, Expressing

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

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.

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: Staining, Cell Counting

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.

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

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.

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: Staining

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.

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: Expressing, Quantitative RT-PCR, Gene Expression

Journal: iScience

Article Title: Proteomics- and BRET- screens identify SPRY2 as a Ras effector that impacts its membrane organization

doi: 10.1016/j.isci.2025.113974

Figure Lengend Snippet: SPRY2 is recruited to active K-Ras on the plasma membrane (A) AlphaFold 3 prediction of K-Ras/SPRY2 complex, with zoom-in showing residues with putative side chain interactions. (B and C) BRET-titration curves of the nL-tagged K-RasG12V or K-RasG12V-Q25A/Y40A/R41A (mutK-RasG12V) interactions with mNG-tagged SPRY2 or SPRY2-K187A/Y191A/T298A (mutSPRY2) (B), respectively corresponding C-SPRY2 variants (C) in HEK cells from N = 2-4 independent biological repeats. Means ± SEM of BRETtop were analyzed using One-Way Brown-Forsythe and Welch ANOVA tests with Dunnett's T3 correction for multiple comparisons. (D) AlphaFold 3 prediction of the SPRY2/SPRY2 interaction, with zoom-in showing residues within 3 Å distance. (E) BRET-titration curves of the nL-SPRY2/mNG-SPRY2 and nL-SPRY2/mNG-SPRY4 interaction in HEK cells with means ± SEM of BRETtop from N = 4 independent biological repeats. (F) BRET-titration curve of the wt or mutant nL-K-RasG12V with mNG-SPRY4 in HEK cells with means ± SEM of BRETtop from N = 3–6 independent biological repeats. (G) Flow cytometric quantification of MyHC terminal differentiation marker expression in C2C12 cells in low serum for 72 h after transfection with mNG-SPRY2, mNG-N-SPRY2, or mNG-C-SPRY2 constructs. Means ± SD are plotted from N = 3 biological repeats. Statistical analysis was done using one-way ANOVA and Dunn’s post hoc test. (H) Schematic illustrating our speculative models for how APLP2 (left) and SPRY2 (right) impact Ras membrane organization and activity. See the main text for more details. See also and .

Article Snippet: The mouse muscle C2C12 cell line (female mouse origin) was purchased from ATCC (CRL-1772) and cultured in DMEM supplemented with ∼9% (v/v) FBS, 2 mM L- and penicillin-streptomycin 10,000 units/mL (high serum medium).

Techniques: Clinical Proteomics, Membrane, Titration, Mutagenesis, Marker, Expressing, Transfection, Construct, Activity Assay