Splicing factor SRSF1 is indispensable for regulatory T cell homeostasis and function

Purpose: The goals of this study are to determine the role of the serine/ arginine-rich splicing factor 1 (SRSF1) in regulatory T cells Overall design: Methods: Total T cells were isolated from spleens of 8-10-week-old WT and T cell Srsf1-KO mice (n=3 each). CD4+CD25+CD127lo natural (n)Treg cells were sorted by flow cytometry. nTregs were stimulated with anti-CD3 (2 µg/ml) and anti-CD28 (2 µg/ml) antibodies and rIL-2 (10 ng/ml) for 72h. Cells were collected, and total RNA was extracted using the RNeasy mini kit (Qiagen) and submitted for RNA sequencing to the Molecular Biology Core Facilities at the Dana-Farber Cancer Institute (DFCI). Libraries were prepared using Illumina TruSeq Stranded mRNA sample preparation kits according to the manufacturer's protocols. Samples were sequenced on an Illumina NextSeq500 run with single-end 75-bp reads. Data analyses were performed using the VIPER pipeline by the DFCI Core.