Igg fitc

A HFD diet (60 kcal %, D12492) and a basal diet (10 kcal %, D12450B) were acquired from Research Diets. Bovine serum, fetal Bovine serum (FBS), and Dulbecco’s modified Eagle’s medium (DMEM) were all purchased from GIBCO (Invitrogen, CA, USA). Insulin, 3-isobutyl-1-metyl-xanthine (IBMX), dexamethasone (DXM), Oil red O working solution, Wortmannin (Wort.) and the primary antibodies for anti-Twist 1 and GluT4 were Sigma products (Merck, German). Primary antibodies for anti-UCP 2, NRF-1, pGC-1 α, pGC-1 β, β-actin, and mtTFA, HRP-conjugated secondary antibodies, and IgG-FITC were all obtained from Abcam. Primary antibodies for anti-p-IRS-1, p-ERK, p-AKT, and p-PI 3 K were CST products. 2-deoxy-D-[1,2-,3 H]-glucose was obtained from Perkin Elmer. The lentiviral vectors pGLV3/H1/GFP + Puro (LV3) and recombinant vectors LV3/Twist 1 shRNA were acquired from Gene Pharma (Shanghai, China). NE-PER^TM Nuclear and Cytoplasmic Extraction Kits, Mem-PER^TM Plus Membrane Protein Kits, and Mitochondria Isolation Kits were all kindly supplied by Pierce (Thermo Scientific). SYBR® Green Real-time PCR Master Mix (QPK-201 T) was purchased from TOYOBO. All other reagents were obtained from Sigma.