Single cell suspensions from spleen and bone marrow were depleted of erythrocytes using ACK lysis buffer and stained with a live/dead marker and the respective antibodies in PBS. Three replicates of each cell population (1,000 cells each) were sorted directly into TRIzol (Thermo Fisher). RNA was purified using the RNeasy Plus Micro kit (QIAgen) according to the manufacturer's instructions. Un-stranded, non-rRNA, non-polyA+ selected libraries were prepared using the SMARTer Ultra Low Input RNA kit for Sequencing v3 (Clontech Laboratories). RNA sequencing: The libraries were sequenced on the Illumina HiSeq 2000 platform as 50 bp paired-end runs
GEO Accession
GSM4697467