Real time fluorescence quantitative polymerase chain reaction

THP-1 cells (from Chinese Academy of Science, Shanghai, China) were cultured in RPMI (SH30809.01B, Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (16000-044, Invitrogen, Carlsbad, CA, USA) at 37 °C. LPS, fetal bovine serum (FBS), phosphate-buffered brine (PBS), polyvinylidene fluoride (PVDF), were purchased from Thermo Fisher Scientific, Waltham, USA. The TRIzol reagent, complementary deoxyribonucleic acid (cDNA) first-strand synthesis kit, and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) detection kit were purchased from TaKaRa, Beijing, China; The cell protein extraction kit and bicinchoninic acid (BCA) protein quantitative detection kit was supplied by KGI Biotech, Jiangsu, China; The methyl thiazolyl tetrazolium (MTT) kit, TLR4, ERK and nuclear factor κB (NF-κB) p65, and β-actin antibodies were ordered from Abcam Trading Co., Ltd., Shanghai, China. CLI-095 and horseradish peroxidase-labeled IgG antibodies were purchased from Millipore, Billerica, MA, USA; An enhanced chemiluminescence (ECL) kit was purchased from Thermo Fisher Scientific, Waltham, USA; And enzyme-linked immunosorbent assay (ELISA) kits for monocyte chemoattractant protein 1 (MCP-1), matrix metalloproteinase 9 (MMP-9), secretion levels of IL-6 and IL-1β were ordered from (Proteintech, Wuhan, China).