RNASeq of RNA from naïve B cells and T cell blasts from individuals with AIOLOS N160S mutation and healthy controls

RNA-seq was performed using RNA extracted from enriched T cell blasts (CD3 T cells were enriched from the T-cell blasts, purity >90%, Stemcell Technologies, 19051) or naïve B cells (purity >90%, Stemcell technologies, 19254) for the following study groups: healthy controls (HC, n=3-4) and patients with AIOLOS N160S (n=1 for naïve B cells [A.III.1] and n=2 for T cell blasts [A.II.2 and A.III.1]). Libraries were prepared using the AmpliSeq for Illumina Transcriptome Human Gene Expression panel, which captures 20,802 genes (>95% of human RefSeq genes). RNA-seq was performed on the Illumina HiSeq 2500 (Illumina; AmpliSeq for Illumina/HiSeq 2500). Demultiplexed reads were mapped to the hg19 genome using the splice-aware aligner Tophat (Trapnell et al., 2009). Gene-level counts data were generated using the Rsubread feature counts a read summarization program that counts mapped reads for genomic features such as genes(Liao et al., 2019). Differential expression analysis was performed using R (v.3.5.3) and DESeq2 (v.1.22.2)(Love et al., 2014). RNA sequencing was performed using the AmpliSeq for Illumina Transcriptome Human Gene Expression panel anf the Illumina HiSeq 2500 analyzer. We analyzed RNA samples extracted from enriched T cell blasts and naïve B cells from healthy controls and patients with AIOLOS N160S mutations.