c2c12  (ATCC)

Journal: Biomedical Reports

Article Title: Both asparagine and proline are required to decrease Sestrin2 mRNA levels via ATF4 reduction and regulate collagen type I alpha 1 chain production and the proliferation of quiescent RI?T hepatic stellate cells

doi: 10.3892/br.2025.1971

Figure Lengend Snippet: Addition of aspartic acid, asparagine, glutamic acid and proline (4AAs) does not influence SESN2 and ATF4 mRNA levels in C2C12, GH3 and HepG2 cells, which exhibit higher ASNS mRNA levels than RI-T cells. (A) C2C12, (B) GH3 and (C) HepG2 cells were treated with 4AAs for 5 h, and total RNA was isolated from the cells. SESN2 and ATF4 mRNA levels were analyzed by RT-qPCR (n=6 per treatment group). Statistical differences were determined using Student's t-test. The addition of 4AAs had no significant effect on SESN2 or ATF4 mRNA levels. (D) ASNS mRNA levels in RI-T, C2C12, HepG2 and GH3 cells were determined by RT-qPCR (n=6 per treatment group). Statistical differences were determined using ANOVA followed by Tukey-Kramer test. # P<0.05 vs. RI-T. ASNS, asparagine synthetase; SESN2, sestrin 2; ATF4, activating transcription factor 4; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: C2C12, HepG2 (a liver cancer cell line; cat. no. RCB1648; Riken Cell Bank) and GH3 (a pituitary cell line producing growth hormone and prolactin; cat. no. CCL-82.1; American Type Culture Collection) cells were cultured in DMEM supplemented with 15.5 µg/ml kanamycin, 100 µg/ml penicillin G and 10% FBS.

Techniques: Isolation, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: The FASEB Journal

Article Title: NOR ‐1 Overexpression Elevates Myoglobin Expression via PERM1 and Enhances Mitochondrial Function and Endurance in Skeletal Muscles of Aged Mice

doi: 10.1096/fj.202500375R

Figure Lengend Snippet: NOR‐1 overexpression elevates metabolic gene expression in C2C12 myotubes. (A) Vector map of AAV9 used for NOR‐1 overexpression. (B) Images showing GFP+ myotubes after AAV9 transduction. The scale bar is 200 μm. (C) NOR‐1 gene expression 7 days after AAV9 transduction ( n = 6). (D) Metabolic gene expression in C2C12 myotubes after NOR‐1 overexpression ( n = 6). (E) Experimental design: C2C12 myotubes were differentiated for 3 days before being transduced with AAVs to express either NOR‐1 or GFP control protein. 5 days after transduction, NOR‐1‐GFP and control‐GFP expressing myotubes were treated with either a PERM1 targeting or negative control siRNA for 2 days before being harvested. Expression of (F) PERM1, (G) CKMT2, and (H) myoglobin after AAV transduction and siRNA transfection ( n = 6). HPRT was used as a housekeeping gene. Data are presented as Mean ± SEM. (C and D) are analyzed using unpaired t ‐test or unpaired t ‐test with Welch's correction when unequal variance was detected. (F–H) were analyzed using One‐Way ANOVA or Brown‐Forsythe and Welch ANOVA when unequal variance was detected. Statistical significance was set to p < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Different letters (A, B, C) denote significant differences between groups. (A) The vector map was created with Vectorbuilder. (E) Made with Biorender.com (Created in BioRender. Paez, H. (2024) https://BioRender.com/g53o551 ).

Article Snippet: Total RNA was isolated from whole muscle or myoblasts derived from C2C12 cells using TRIzol reagent (Invitrogen) per manufacturer instructions.

Techniques: Over Expression, Gene Expression, Plasmid Preparation, Transduction, Control, Expressing, Negative Control, Transfection

Journal: The FASEB Journal

Article Title: NOR ‐1 Overexpression Elevates Myoglobin Expression via PERM1 and Enhances Mitochondrial Function and Endurance in Skeletal Muscles of Aged Mice

doi: 10.1096/fj.202500375R

Figure Lengend Snippet: NOR‐1 overexpression elevates metabolic gene expression in C2C12 myotubes. (A) Vector map of AAV9 used for NOR‐1 overexpression. (B) Images showing GFP+ myotubes after AAV9 transduction. The scale bar is 200 μm. (C) NOR‐1 gene expression 7 days after AAV9 transduction ( n = 6). (D) Metabolic gene expression in C2C12 myotubes after NOR‐1 overexpression ( n = 6). (E) Experimental design: C2C12 myotubes were differentiated for 3 days before being transduced with AAVs to express either NOR‐1 or GFP control protein. 5 days after transduction, NOR‐1‐GFP and control‐GFP expressing myotubes were treated with either a PERM1 targeting or negative control siRNA for 2 days before being harvested. Expression of (F) PERM1, (G) CKMT2, and (H) myoglobin after AAV transduction and siRNA transfection ( n = 6). HPRT was used as a housekeeping gene. Data are presented as Mean ± SEM. (C and D) are analyzed using unpaired t ‐test or unpaired t ‐test with Welch's correction when unequal variance was detected. (F–H) were analyzed using One‐Way ANOVA or Brown‐Forsythe and Welch ANOVA when unequal variance was detected. Statistical significance was set to p < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Different letters (A, B, C) denote significant differences between groups. (A) The vector map was created with Vectorbuilder. (E) Made with Biorender.com (Created in BioRender. Paez, H. (2024) https://BioRender.com/g53o551 ).

Article Snippet: To induce differentiation, C2C12 myoblasts were grown to 70% confluence and GM was replaced with differentiation media (DM) containing DMEM supplemented with 2% heat‐inactivated horse serum (Gibco) and 1% antibiotic/antimycotic.

Techniques: Over Expression, Gene Expression, Plasmid Preparation, Transduction, Control, Expressing, Negative Control, Transfection

Journal: PLOS One

Article Title: The potential function of soy protein hydrolysate to induce myogenic differentiation of C2C12 cells

doi: 10.1371/journal.pone.0321650

Figure Lengend Snippet: C2C12 cells were cultured for 24 h in the presence of different concentrations of FBS and 0.5 g/L different types of HVPs.

Article Snippet: To find a serum substitute for C2C12 cells, we screened a variety of Angel Yeast-produced hydrolyzed vegetable protein (HVP), using one SPH product from Becton, Dickinson Company as a reference.

Techniques: Cell Culture

Journal: PLOS One

Article Title: The potential function of soy protein hydrolysate to induce myogenic differentiation of C2C12 cells

doi: 10.1371/journal.pone.0321650

Figure Lengend Snippet: The cell viability was detected by CCK-8 reagents after C2C12 cells culturing in different conditions for 24 h(A) and 48 h(B), and 10%FBS group was used as a standard to process data. * represents p < 0.05, and *** represents p < 0.001.

Article Snippet: To find a serum substitute for C2C12 cells, we screened a variety of Angel Yeast-produced hydrolyzed vegetable protein (HVP), using one SPH product from Becton, Dickinson Company as a reference.

Techniques: CCK-8 Assay

Journal: PLOS One

Article Title: The potential function of soy protein hydrolysate to induce myogenic differentiation of C2C12 cells

doi: 10.1371/journal.pone.0321650

Figure Lengend Snippet: A: MyHC was detected by TRITC red fluorescence after immunofluorescence after C2C12 cells cultured in different conditions for 24 h; B-C: The MFI of MyHC marked with TRITC red fluorescence after immunofluorescence were quantitatively measurd and analyzed. * represents p < 0.05, ** represents p < 0.01 and *** represents p < 0.001.

Article Snippet: To find a serum substitute for C2C12 cells, we screened a variety of Angel Yeast-produced hydrolyzed vegetable protein (HVP), using one SPH product from Becton, Dickinson Company as a reference.

Techniques: Fluorescence, Immunofluorescence, Cell Culture

Journal: PLOS One

Article Title: The potential function of soy protein hydrolysate to induce myogenic differentiation of C2C12 cells

doi: 10.1371/journal.pone.0321650

Figure Lengend Snippet: The expression levels of MyoD, MyHC and MyoG were analyzed by qPCR in C2C12 cells cultured in different conditions for 2 days(A) and 5 days(B). 10%FBS group was used as a standard to process data. ns represents no significance, and *** represents p < 0.001.

Article Snippet: To find a serum substitute for C2C12 cells, we screened a variety of Angel Yeast-produced hydrolyzed vegetable protein (HVP), using one SPH product from Becton, Dickinson Company as a reference.

Techniques: Expressing, Cell Culture

Journal: The FASEB Journal

Article Title: NOR ‐1 Overexpression Elevates Myoglobin Expression via PERM1 and Enhances Mitochondrial Function and Endurance in Skeletal Muscles of Aged Mice

doi: 10.1096/fj.202500375R

Figure Lengend Snippet: NOR‐1 overexpression elevates metabolic gene expression in C2C12 myotubes. (A) Vector map of AAV9 used for NOR‐1 overexpression. (B) Images showing GFP+ myotubes after AAV9 transduction. The scale bar is 200 μm. (C) NOR‐1 gene expression 7 days after AAV9 transduction ( n = 6). (D) Metabolic gene expression in C2C12 myotubes after NOR‐1 overexpression ( n = 6). (E) Experimental design: C2C12 myotubes were differentiated for 3 days before being transduced with AAVs to express either NOR‐1 or GFP control protein. 5 days after transduction, NOR‐1‐GFP and control‐GFP expressing myotubes were treated with either a PERM1 targeting or negative control siRNA for 2 days before being harvested. Expression of (F) PERM1, (G) CKMT2, and (H) myoglobin after AAV transduction and siRNA transfection ( n = 6). HPRT was used as a housekeeping gene. Data are presented as Mean ± SEM. (C and D) are analyzed using unpaired t ‐test or unpaired t ‐test with Welch's correction when unequal variance was detected. (F–H) were analyzed using One‐Way ANOVA or Brown‐Forsythe and Welch ANOVA when unequal variance was detected. Statistical significance was set to p < 0.05. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Different letters (A, B, C) denote significant differences between groups. (A) The vector map was created with Vectorbuilder. (E) Made with Biorender.com (Created in BioRender. Paez, H. (2024) https://BioRender.com/g53o551 ).

Article Snippet: C2C12 cells were purchased from ATCC (Manassas, VA) and propagated in growth media (GM) consisting of Dulbecco's modified Eagle's medium (DMEM; Invitrogen) with 10% fetal bovine serum (FBS; Gibco) and supplemented with 1% antibiotic/antimycotic (Gibco).

Techniques: Over Expression, Gene Expression, Plasmid Preparation, Transduction, Control, Expressing, Negative Control, Transfection

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Transcriptional Intermediary Factor 1γ–Induced Irisin in Skeletal Muscle Attenuates Renal Fibrosis in Diabetic Nephropathy

doi: 10.1002/jcsm.13810

Figure Lengend Snippet: Effect of TIF1γ on irisin expression and secretion in C2C12 cells. (A & B) Experimental in vitro scheme for C2C12 differentiation. (C & D) Changes in irisin expression and secretion following TIF1γ plasmid (2 μg) transfection during C2C12 differentiation. Statistical significance was determined using a one‐way analysis of variance followed by Tukey's multiple comparison test. Data are presented as mean ± SEM. * p < 0.05. TIF1γ, transcriptional intermediary factor 1γ; C2C12, murine myoblast cells; SEM, standard error of the mean.

Article Snippet: The C2C12 mouse skeletal muscle myoblasts were purchased from the American Type Culture (ATCC, USA).

Techniques: Expressing, In Vitro, Plasmid Preparation, Transfection, Comparison

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Transcriptional Intermediary Factor 1γ–Induced Irisin in Skeletal Muscle Attenuates Renal Fibrosis in Diabetic Nephropathy

doi: 10.1002/jcsm.13810

Figure Lengend Snippet: Effect of TIF1γ on palmitate treatment in C2C12 cells and the paracrine effect of TIF1γ‐induced irisin on palmitate‐treated HK‐2 cells. (A) Changes in irisin, TIF1γ, PGC‐1α and pAkt expression following palmitate (100 μM) treatment in the presence or absence of TIF1γ (2 μg). (B) Changes in irisin secretion following palmitate (100 μM) treatment with or without TIF1γ (2 μg). (C) Changes in EMT markers in palmitate (100 μM)‐treated HK‐2 cells induced by CM from TIF1γ‐transfected C2C12 cells. (D) Western blot analysis of FNDC5 protein levels in C2C12 cells transfected with control or FNDC5‐specific siRNAs (#1–5) for 48 h. (E) Irisin levels in CM from C2C12 cells transfected with 25 nmol siRNA‐FNDC5 and 2 μg TIF1γ, measured by ELISA. (F) Changes in EMT markers in palmitate (100 μM)‐treated HK‐2 cells induced by CM from C2C12 cells transfected with 25 nmol siRNA‐FNDC5 and TIF1γ. Each protein expression level was normalized to α‐tubulin or β‐actin (A, C, D, F). TIF1γ, transcriptional intermediary factor 1γ; HK‐2, human kidney 2 cells; C2C12, murine myoblast cells; PGC‐1α, peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha; pAkt, phosphorylated protein kinase B; EMT, epithelial–mesenchymal transition; BMP‐7, bone morphogenetic protein 7; α‐SMA, alpha‐smooth muscle actin; pSmad2/3, phosphorylated Smad2/3; CM, conditioned medium; siRNA, signal interfering RNA.

Article Snippet: The C2C12 mouse skeletal muscle myoblasts were purchased from the American Type Culture (ATCC, USA).

Techniques: Expressing, Transfection, Western Blot, Control, Enzyme-linked Immunosorbent Assay