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.