GSM4759442: NKT17_lung_A; Mus musculus; RNA-Seq
Cells representing specific iNKT subsets were sorted directly into 0.2 ml PCR tubes containing 8 µL of low-input lysis buffer (0.2% Triton-X-100 and RNase inhibitor) and stored at -80oC until processed further. RNA extraction, reverse transcription, cDNA amplification, fragmentation, and additional PCR amplification were performed as previously described (Rosales et al. Methods Mol Biol 1799, p275). We followed the Smart-Seq2 protocol as described previously (Picelli et al., 2013; Rosales et al., 2018). Libraries were then pooled at equal molar concentration and sequenced using the HiSeq 2500 and NovaSeq6000 Illumina platform (Illumina). Samples from the experiment comparing splenic NKTFH and NKTeff cells from aGalCer-challenged mice with unchallenged splenic iNKT cells were sequenced via a paired-end read method using the Novaseq6000 sequencer according to supplied protocols (Illumina). Sequencing for all other samples was performed according to a single-end strategy using the HiSeq2500 sequencer (Illumina).
GEO Accession
GSM4759442