RNA-Seq gene profiling comparison from human cDC subsets and pre-cDC subsets

The goal of this study is to compare the gene profiling of human blood BDCA-1+ cDCs, blood BDCA-3+ cDCs, CB CD172a-pre-cDCs and CB CB172a+ pre-cDCs. RNA from sorted populations was extracted and column-purified using the Arcturus PicoPure RNA Isolation kit (Applied Biosystems). Genomic DNA was removed by on-column digest with DNAse I (Qiagen), according to the PicoPure RNA Isolation kit manual. RNA libraries were prepared using the SMARTer Ultra Low Input RNA for Sequencing kit (Clontech Laboratories) followed with Nextera XT DNA Library Prep kit (Illumina). Libraries were sequenced by 75-bp single-end reading on a NextSeq 500 sequencer (Illumina). Our results show that CD172a+ and CD172a- pre-cDCs represent developmentally discrete populations that differentially express lineage-restricted transcription factors. Moreover CD172a- pre-cDCs cluster with BDCA-3hi cDCs and CD172a+ pre-cDCs with BDCA-1+ cDCs. RNA from sorted cord blood pre-cDC subsets (150-400 cells) and peripheral blood cDC subsets (2000-10000 cells). Done in quadruplicate.