cell culture c2c12 mouse myoblasts  (ATCC)

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: RORα-GABP-TFAM axis alleviates myosteatosis with fatty atrophy through reinforcement of mitochondrial capacity.

doi: 10.1002/jcsm.13432

Figure Lengend Snippet: Figure 6 Activation of retinoic acid receptor-related orphan receptor-α (RORα) restored the mitochondrial function in glycolytic fibres. (A) Mouse gas- trocnemius (GA) tissues were subjected to immunostaining for MyHC2b (red). Succinate dehydrogenase (SDH) staining was performed in the GA sec- tions of the high-fat diet (HFD)-fed and AAV-RORα-treated mouse model. Tissues were examined by using an automated multimodal tissue analysis system, and representative images are shown. Scale bar: 1 mm (MyHC2b and SDH activity) or 50 μm (RORα). (B) C2C12 cells were exposed to differ- entiation medium containing 0.1-mM palmitic acid (PA) conjugated with bovine serum albumin (BSA) and treated with cholesterol sulfate (CH-sulfate) or JC1-40. After 7 days, total RNA was isolated and mRNA level of Myh4 was measured by quantitative real-time PCR (qRT-PCR). ***P < 0.001 versus vehicle (n = 4). (C) Immunostaining was performed with C2C12 cells in the presence of 0.1-mM PA conjugated with BSA at Day 7 of differentiation for staining of MyHC2b (red) and DAPI (blue). Representative images examined by confocal microscopy are shown. Data were obtained from three inde- pendent experiments, and fluorescence intensity was quantified in 10 images of each group by using ImageJ. Scale bar: 25 μm. ***P < 0.001 versus vehicle.

Article Snippet: Cell lines and cell culture Mouse myoblast C2C12 cells (CRL-1772, ATCC, Manassas, VA, USA) were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Hyclone, Logan, UT, USA) containing 10% foetal bovine serum (FBS).

Techniques: Activation Assay, Immunostaining, Staining, Activity Assay, Isolation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Confocal Microscopy