Ficoll lymphocyte separation medium

THP-1 human macrophages were purchased from Shanghai Institute of Cell Research, Chinese Academy of Sciences; Mtb standard strain H37Rv (ATCC 27294) was preserved by the laboratory. The Ficoll lymphocyte separation medium, fetal bovine serum, and phorbol myristate acetate (PMA) were procured from Sigma (USA); the transfection reagent Lipofectmine 2000 and Trizol from Invitrogen (USA); the PrimeScript reverse transcription reagent kit with gDNA Eraser and SYBR Premix Ex Taq from TaKaRa (Dalian, China); DMEM medium from Hyclone (USA); the Opti-MEM I Reduced Serum Medium for transfection from Gibco (USA); the Middlebrook 7H10 medium and OADC from BD (USA); and the enzyme-linked immunosorbent assay (ELISA) kit detecting human TNF-α and IL-6 from eBioscience (USA). The spectrophotometer was purchased from Bio-Rad (USA), and the Applied Biosystems 7600 thermocycler from ABI (USA).
PBMC Isolation. Morning fasting venous blood was collected in a 5-mL tube with heparin sodium for anticoagulation. PBMCs were routinely isolated using the Ficoll lymphocyte separation medium.

Mtb standard strain H37Rv (ATCC 27294) was preserved by the laboratory. THP-1 human macrophages were purchased from the Shanghai Institute of Cell Research, Chinese Academy of Sciences. Phorbol-12-myristate-13-acetate (PMA) and Ficoll lymphocyte separation medium were purchased from Sigma (USA). The RPMI-1640 medium, fetal bovine serum, and phosphate-buffered saline (PBS) were obtained from BI (Israel). Trizol was purchased from Invitrogen (USA). Chloroform, isopropanol, and ethanol were purchased from Damao Chemical Reagent Factory (China). DNase/RNase-free double-distilled water and RNAwait were obtained from Solarbio (China). The PrimeScript reverse transcription reagent kit with gDNA Eraser and TB Green Premix Ex Taq II were purchased from TaKaRa (Japan).