GSM4679357: ADAM10 KO CD4+ DC_sample 1; Mus musculus; RNA-Seq
Spleens were cut in grain size pieces and digested with 200U/ml Collagenase Type IV + 0,5 U/ml Dnase for 30 min at 37 oC. CD11c+ cells were pre-enriched by MACS, followed by FACS-sorting for living CD11c+MHCII+CD8+ cDC1, CD11c+MHCII+CD4+ cDC2 and CD11c+MHCII+CD8-CD4- cDC2. mRNA was purified from sorted splenic cDC subpopulations with the RNeasy Plus Micro kit according to manufacturer's instructions (Qiagen). Barcoded mRNA-seq cDNA libraries were prepared from 50 ng of total mRNA using NEBNext Poly(A) mRNA Magnetic Isolation Module and NEBNext Ultra II RNA Library Prep Kit for Illumina according to the manual (New England BioLabs. Barcoded RNA-Seq libraries were onboard clustered using HiSeq Rapid SR Cluster Kit v2 using 8 pM and 59 bps reads were sequenced on the Illumina HiSeq2500 using the HiSeq Rapid SBS Kit v2 (50 Cycles).
GEO Accession
GSM4679357