GSM5509772: CD4_CD103–TRM_Stim_D07_MildAsthma_S2; Homo sapiens; RNA-Seq
For bulk RNA-seq, a maximum of 400 cells per population of interest, per donor, and per stimulation condition were sorted individually into low retention 0.2mL PCR tubes, containing 8uL of ice-cold lysis buffer (Triton X-100 [0.1%, Sigma-Aldrich], 1% RNase inhibitor [Takara Bio]). Following sorting, tubes were vortexed for 10s, span for 1min at 3000g, and directly stored at -80°C. Bulk RNA-seq libraries were prepared for sequencing using the Smart-seq2 protocol adapted for samples with small cell numbers (Rosales et al, 2018) and following standard Illumina protocols. 100 x 100 bp paired-end reads,  Hg19 human reference genome.