MicroRNA profiling in interleukin-27 yreated monocyte-derived human dendritic cells
CD14+ monocytes were isolated from three donors: DC2, DC4, DC7. Monocyte-derived immature DCs (iDCs) were generated by incubating 0.5x10^6/ml monocytes in G4 medium at 37 degree in a CO2 (5%) incubator for seven days. The IL-27 treated DCs (27DCs) were induced from monocytes cultured in G4 medium (iDCs (1x10^6) stimulated) with IL-27 (100 ng/mL) at 37 degree for seven days. The culture media was changed with fresh G4 media every 3–4 days. Total six samples were in this study. Three controls are iDC_2, iDC_4, iDC_7; Three IL27-treated samples are 27DC_2, 27DC_4, 27DC_7.   Total RNA was extracted with chloroform and phenol. Prior to RNA sequencing, samples were subjected to analysis by the Agilent Bioanalyzer RNA Nano chip and the small RNA chip to confirm RNA purity and quality (Santa Clara, CA, USA). All samples had an RNA integrity number of 10. Six microRNA libraries were prepared using the NEBNext Multiplex Small RNA Library Prep for Illumina protocol (New England BioLabs, Ipswich, MA, USA) according to the manufacturer’s instructions and sequenced on the Hiseq 2500 using v4 chemistry for single end sequencing (San Diego, CA, USA).