GSM8942730: ILC1/3Ps-1 from fetal liver of WT mice; Mus musculus; RNA-Seq
To obtain CLP (Lin–CD45+CD127+Flt3+CD25– live cells), ILCP (Lin–CD45+CD127+Flt3–α4β7+CD25–PD-1+CD94– live cells) and ILC1/3P (Lin–CD45+CD127+Flt3–α4β7+CD25–PD-1–CD94+ live cells) were sorted from the fetal liver of WT B6 mice. Mononuclear cells (MNCs) were incubated with anti-CD45 mAbs for 30 min at 4°C. Subsequently, MNCs were incubated with anti-PE beads for 60 min at 4°C. Nextly, MNCs were incubated with the indicated antibodies for 30 min at 4°C. 7AAD was added to the MNCs suspension, which was subsequently sorted using a FACSAria Fusion (BD Biosciences, San Diego, USA). The purity of the sorted cell populations was >  90%. Purified live cells were labelled with 0.4% Trypan blue to determine their viability (> 90% before processing for RNA-seq). Purified cells were gently transferred into lysis buffer, and reverse transcription was performed using a SMARTer Ultra Low RNA Kit (Clontech) directly in the cell lysate. Single-cell cDNA was amplified using an Advantage 2 PCR Kit (Clontech) according to the manufacturer's protocol. cDNA library was sequenced using the NovaSeq6000.