Journal: Burns & Trauma

Article Title: Enhancing diabetic muscle repair through W-GA nanodots: a nanomedicinal approach to ameliorate myopathy in type 2 diabetes

doi: 10.1093/burnst/tkae059

Figure Lengend Snippet: Evaluation of the biocompatibility of W-GA in vitro. ( a ) C2C12 cells cultured for 7 days in W-GA were evaluated for proliferation and cytotoxicity using live/dead staining. Scale bar = 500 μm. ( b ) Survival rate of C212 cells ( n = 3) is shown as the mean ± standard deviation. The statistical significance of differences between treatments was determined by one-way ANOVA and the Bonferroni posthoc correction. NS: Not significant. ( c ) C2C12 cell proliferation capacity was assessed 24 hours post-treatment using BrdU incorporation. The green signal represents BrdU. Scale bar = 500 μm. ( d ) Quantification of BrdU assay data ( n = 3). The data are presented as the mean ± standard deviation. NS: Not significant. ( e ) The cell proliferation ability of C2C12 cells in the W-GA group was further evaluated using a CCK-8 assay. The data are presented as the mean ± standard deviation. NS: Not significant

Article Snippet: C2C12 myoblasts (a mouse cell line) were obtained from ScienCell Research Laboratories and cultured in Dulbecco's modified Eagle’s medium (DMEM; containing 25 mM glucose; Gibco, USA), which included 10% fetal bovine serum (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco, USA), in a humidified atmosphere at 37°C and 5% CO 2 .

Techniques: In Vitro, Cell Culture, Staining, Standard Deviation, BrdU Incorporation Assay, BrdU Staining, CCK-8 Assay

Journal: Burns & Trauma

Article Title: Enhancing diabetic muscle repair through W-GA nanodots: a nanomedicinal approach to ameliorate myopathy in type 2 diabetes

doi: 10.1093/burnst/tkae059

Figure Lengend Snippet: Antiapoptotic, antioxidative, and myogenic differentiation-promoting effects of W-GA. ( a ) Flow cytometry profiles showing the abundance of total C2C12 cells under various treatment conditions, along with apoptosis events in C2C12 cells under different therapeutic interventions. ( b ) Quantification of flow cytometry data for apoptotic cells ( n = 3). The data are presented as the mean ± standard deviation. NS: Not significant, * * p < 0.01. The statistical significance of differences between treatments was determined by one-way ANOVA and the Bonferroni posthoc correction. ( c ) Flow cytometry profiles showing the production of ROS in C2C12 cells under different treatment conditions. ( d ) Quantification of flow cytometry data for ROS production ( n = 3). The data are presented as the mean ± standard deviation. NS: Not significant, * p < 0.05. ( e ) Representative immunofluorescence image illustrating MYHC and MyoD protein expression in C2C12 myoblasts. Scale bar = 100 μm. ( f ) Quantitative analysis and intergroup comparison of myotube diameters ( n = 3). The statistical significance of differences between treatments was determined by one-way ANOVA and the Bonferroni posthoc correction. NS: Not significant, * p < 0.05, * * p < 0.01

Article Snippet: C2C12 myoblasts (a mouse cell line) were obtained from ScienCell Research Laboratories and cultured in Dulbecco's modified Eagle’s medium (DMEM; containing 25 mM glucose; Gibco, USA), which included 10% fetal bovine serum (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco, USA), in a humidified atmosphere at 37°C and 5% CO 2 .

Techniques: Flow Cytometry, Standard Deviation, Immunofluorescence, Expressing, Comparison

Journal: Journal of Biomedical Science

Article Title: Vitamin D3/VDR resists diet-induced obesity by modulating UCP3 expression in muscles

doi: 10.1186/s12929-016-0271-2

Figure Lengend Snippet: Calcitriol enhanced relative luciferase activity in a dose dependent manner. UCP3-pro-luc was transfected into C2C12 cells ( a ), L6 cells ( b ), and H-EMC-SS cells ( c ). Relative luciferase activity was analyzed by Dual Luciferase Assay after stimulation with calcitriol. n = 3, compared with vehicle, *: P < 0.05; **: P < 0.01

Article Snippet: Mouse C2C12 muscle cells (C2C12), rat L6 muscle cells (L6), H-EMC-SS chondrosarcoma cells, and human embryonic kidney cells (HEK293) were purchased from JCRB Cell Bank (National Institute of Biomedical Innovation, Tokyo, Japan).

Techniques: Luciferase, Activity Assay, Transfection

Journal: Journal of Biomedical Science

Article Title: Vitamin D3/VDR resists diet-induced obesity by modulating UCP3 expression in muscles

doi: 10.1186/s12929-016-0271-2

Figure Lengend Snippet: Calcitriol increased UCP3 mRNA expression in muscle cells. Relative mRNA expression was detected in C2C12 cells ( a ), L6 cells ( b ), and H-EMC-SS cells ( c ) using qRT-PCR. To investigate the role of calcitriol and VDR on UCP3 mRNA expression, we transfected with a control plasmid ((-) mock) or VDR expression plasmid ((+) pCMX-VDR). We also performed loss-of-function experiments using selective siRNA against UCP3 ( a ). Compared with non-transfected controls, siRNAs against UCP3 strongly reduced the expression of the UCP3. n = 3. Compared with vehicle, *; P < 0.05, **; P < 0.01. ***; P < 0.005

Article Snippet: Mouse C2C12 muscle cells (C2C12), rat L6 muscle cells (L6), H-EMC-SS chondrosarcoma cells, and human embryonic kidney cells (HEK293) were purchased from JCRB Cell Bank (National Institute of Biomedical Innovation, Tokyo, Japan).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: CRIF1 gene therapy ameliorates inflammatory bowel disease by suppressing TH17 cells and fibrosis through mitochondrial function regulation

doi: 10.3389/fimmu.2025.1618012

Figure Lengend Snippet: Transplantation of CRIF1-overexpressing mitochondria alleviates DSS-induced colitis. (A) Chronic DSS-induced colitis was induced through oral administration of 3% DSS in distilled water for 5 days, followed by regular drinking water three times as shown in the schematic. Mitochondria were isolated from C2C12 cells transfected with p3XFLAG-CMV-10-CRIF1 or mock vector and mitochondria (10 μg in 200 μL saline) were intraperitoneally injected 1 day before and on days 2, 4, 6, 9, 12, and 14 from the start of treatment (n = 5/group). (B) Percentage change in body weight during the disease process. (C) On day 38 after the first DSS administration, colon sections were stained with hematoxylin and eosin (H&E) and antibodies against CRIF1, α-smooth muscle actin (α-SMA), and collagen-I (Col-I). Representative images are shown (original magnification: H&E 200×; IHC 400×, scale bar: 100 µm). Graphs show the mean histological scores for H&E and the numbers of antibody-positive cells for IHC. (D) On day 38 after the first DSS administration, the frequencies of IL-17+CD4+ and CD5+CD1d+IL-10+CD19+ cells in ex vivo spleens of mice were analyzed via flow cytometry. (E) CCD-18Co cells were transfected with pCMV6-Myc-DDK hCRIF1 vector or mock vector; 3 days later, mitochondria were isolated from these cells. Mitochondria (0.3 μg/mL) were cocultured with human PBMCs under the stimulation of anti-CD3 antibody (2 μg/mL); 3 days later, the supernatant was harvested and the amount of IL-17 was measured by ELISA. Values are presented as means ± SDs. *P < 0.05, **P < 0.01. Data are representative of two independent experiments.

Article Snippet: C2C12 cells cells (Korean cell line bank, Seoul, Korea), which are myoblasts from mouse muscle, were transfected with p3XFLAG-CMV-10-CRIF1 or mock vector using Lipofectamine, in accordance with the manufacturer’s recommendation.

Techniques: Transplantation Assay, Isolation, Transfection, Plasmid Preparation, Saline, Injection, Staining, Ex Vivo, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Research

Article Title: Doping-Engineered Proangiogenic Nanozymes Orchestrate Ischemic Tissue Regeneration via Cytoprotection and Revascularization

doi: 10.34133/research.1260

Figure Lengend Snippet: Cu-doped Prussian blue (CuPB) nanozymes protect C2C12 myoblasts and H9c2 cardiomyocytes from H 2 O 2 -induced oxidative injury. (A and B) Representative fluorescence images and quantification of intracellular reactive oxygen species (ROS) in H 2 O 2 -injured C2C12 cells after Prussian blue (PB) or CuPB treatment, detected using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe. Scale bar: 50 μm. n = 5. (C and D) Representative terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining images and quantification of apoptotic C2C12 cells following H 2 O 2 injury with PB or CuPB treatment. Scale bar: 50 μm. n = 5. (E) Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of apoptosis-related genes ( Bcl2 , Caspase3 , Caspase9 , and Bax ) in C2C12 cells after different treatments. n = 3. (F and G) Representative fluorescence images and quantification of intracellular ROS in H 2 O 2 -injured H9c2 cells after PB or CuPB treatment, detected using the DCFH-DA probe. Scale bar: 50 μm. n = 5. (H and I) Representative TUNEL staining images and quantification of apoptotic H9c2 cells following H 2 O 2 injury with PB or CuPB treatment. Scale bar: 50 μm. n = 5. (J) qRT-PCR analysis of apoptosis-related gene expression ( Bcl2 , Caspase3 , Caspase9 , and Bax ) in H9c2 cells after different treatments. n = 5.

Article Snippet: The rat cardiomyocyte cell line (H9c2) was obtained from Procell Life Science & Technology Co., Ltd. (China), and the mouse myoblast cell line (C2C12) was purchased from Beijing Zhongyuan Heju Biotechnology Co., Ltd., the authorized American Type Culture Collection distributor in China (CRL1772).

Techniques: Fluorescence, End Labeling, TUNEL Assay, Staining, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Gene Expression