Fluorescence quantification kit

OPCs (CAS#: 4852-22-6) extracted from grape seeds were provided by Shanghai Yuanye (China). LPS and dexamethasone (DEX) were provided by Macleans (China). MAC-T cells were purchased from BMCC (China). Nuclear protein extraction kits and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazole bromide (MTT) were purchased from Solarbio (China). Fluorescence quantification kits were provided by Takara (China). Primary antibodies against phosphorylated p65 (p-p65), p-ERK1/2 p-IκBα, p65, IκBα and ERK1/2 were obtained from Bioss (China), and antibodies against JNK1/2, p38, p-JNK1/2, and p-p38 were from Wanleibio (China). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (China). NF-κB Activation Nuclear Transport Test Kit and ELISA Kit were supplied by Bevotime (China) and Sinobestbio (China), respectively. Tris-buffered saline + Tween 20 (TBST) was supplied by Solarbio.

ISL and dexamethasone (DEX) were provided by Macleans (Shanghai, China). LPS from Escherichia coli 055:B5 serotype, purity ≥ 99%, item number: L8880) and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazole bromide] were provided by Solarbio (Beijing, China). The fluorescence quantification kits were purchased from Takara (Beijing, China). Primary antibodies were obtained commercially, including p-p65, p-IκBα, p65, IκBα, p-ERK1/2 and ERK1/2 (Bioss, Beijing, China) as well as JNK, p38, p-JNK and p-p38 (Wanleibio, Shenyang, China). Fetal bovine serum (FBS) and Dulbecco's modified Eagle's medium (DMEM) were supplied by Gibco (Suzhou, China). The ELISA and NF-κB Activation Nuclear Transport Test Kits were available from SinoBestBio (Shanghai, China) and Beyotime (Shanghai, China), respectively. Tris-buffered saline plus Tween 20 (TBST) was obtained from Solarbio (Shanghai, China).

Total RNA was extracted from liver and adipose tissue using Trizol reagent (Takara Bio, Shiga, Japan), and RNA concentration was measured using a microspectrophotometer (ThermoFisher, Waltham, MA, USA). One ug of RNA was reverse-transcribed to cDNA using a reverse transcription kit (Takara Bio, Shiga, Japan), and quantitative fluorescence analysis was performed using a fluorescence quantification kit (Takara Bio, Shiga, Japan). The primer sequences are shown in Table 1.