GSM8547183: PP cDC2s+LysoDCs/Blood CD115hi cells; Mus musculus; RNA-Seq
Peyer's patches were digested for 40 min at room temperature in RPMI cell culture medium containing collagenase II/DNase I . All subsequent procedures were at 0–4°C. CD11c+ cells were enriched using anti-CD11c microbeads and an AutoMACS magnetic cell separator according to manufacturer's instructions. CD11c+ cells were preincubated on ice for 10 min with 2.4G2 antibody to block Fc receptors and stained for the  surface markers CD11c, MHCII, SIRPa, MerTK, CD4, embigin, JAM-A, CD24, CD11b, XCR1 and CD101. Cell viability was evaluated using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit (Life Technologies). Live cDC2s and LysoDC subsets were tagged with HTOs either before (second dataset) or after (first dataset) sorting using a  FACSAria III. Blood cells were stained for CD45, CD115, Ly6C, CD43, SIRPa, CD11b, CD11c, MHC-II, CD24 and CCR2. They were tagged with HTOs either before (second dataset) or after (first dataset) sorting using a  FACSAria III. All sorted cells were processed according to the ChromiumTM Single Cell 3' Reagent. Guidelines from 10X genomics. RNA libraries were prepared using the Chromium Single Cell 3' kit protocol from 10X genomics (California, USA)