RNA-Seq
PF Th17 cell subpopulations were sorted from 5 patients and pooled for
RNA-Seq. Cellular RNA was extracted using the Arcturus PicoPure
RNA Isolation Kit (ThermoFisher). The quality and quantities of RNA
samples were measured with the QIAxcel RNA QC Kit v2.0 (Qiagen)
following the manufacturer’s instructions. One microgram RNA at the
concentration > 200 ng/µl was subjected to library
construction, sequencing, reads quality check, alignment, clustering,
expression assessment, Gene Ontology (GO) enrichment analysis, and Kyoto
Encyclopedia of Genes and Genomes (KEGG) enrichment analysis by the
Wuhan SeqHealth Co., Ltd. The CollibriTM Stranded RNA
Library Prep Kit for Illumina (ThermoFisher), HiSeq 2000 Sequencing
System (Illumina), TopHat v2.1.1, HTSeq v0.12.4, DEGseq2, and
Hypergeometric test were used for the above procedures, respectively.