2.7 Cell transfection and culture
To investigate the role of miR-24 and miR-27 in Th2 cells, we performed miR-24 and miR-27 transfection experiments in PBMCs. PBMCs were isolated by Ficoll density gradient. After isolation, Ctrl-m, miR-24-m, or miR-27-m, Ctrl-i, miR-24-i, or miR-27-i (obtained from RiboBio; Guangzhou, China) were transfected into the PBMCs using a riboFECT CP Transfection Kit (Guangzhou, China). Cells were cultured in a 24-well plate at 3x106 cells per ml of RPMI-1640 medium (Gibco, CA, USA) supplemented with 15% fetal calf serum (Gibco, CA, USA) and maintained at 37℃ with 5% CO2. Finally, cells were harvested for flow cytometry and RT-PCR analysis.