Transcriptional profiling of human in vitro primed Th9 T cells (day 20) treated with GW9662 against untreated control
Transcriptional profiling of human in vitro primed Th9 T cells (day 20) treated with GW9662 against untreated control
Human naive T cells were isolated from PBMCs using the EasySep™ Human naive CD4+ T Cell Isolation Kit (Stemcell Technologies) as per the manufacturer's instructions. Culture medium consisted of PRMI-1640 with Hepes (GIBCO) supplemented with 5% of heat-inactivated human serum (Swiss Red Cross, Basel, Switzerland), 2mM L-Glutamine (Biochrom), 50U/ml penicillin and 50μg/ml streptomycin (Bioconcept) and 50 IU/ml IL-2 (Roche). T cells were cultured at a density of 0.25-1x105cells/96 well plate in a total volume of 200μL cell culture medium. Th9 cells were incubated with GW9662 for 48 hours. Total RNA was isolated using the RNAeasy kit from Qiagen. Illumina TruSeq Stranded mRNA protocol (Directional, first strand)