GSM6858324: Control; 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, Bst2, CD4, CD11b, XCR1 and CD24. Cell viability was evaluated using LIVE/DEAD™ Fixable Blue Dead Cell Stain Kit (Life Technologies). Live cDC1s, cDC2s, LysoDC and macrophages were sorted using a FACSAria III and processed according to the ChromiumTM Single Cell 3' Reagent Guidelines from 10X genomics. RNA library was prepared using the Chromium Single Cell 3' kit protocol from 10X genomics (California, USA)