GSM4399494: YUMMER1.7 H2B-eGFP treated with IL-18 [YRWT18]; Mus musculus; RNA-Seq
Tumors were removed from mice on day 13, dissociated by sterile mincing and treated with 1mg/mL collagenase (Sigma) and DNase (1:1000) and 2% FBS in RPMI and incubated for 30 minutes at 37'C, then filtered through a 100 uM cell strainer. Cells were washed, treated with ACK buffer to remove red blood cells and washed again. To sort, cells were stained wtih anti-CD45 (BioLegend; 1:100), anti-CD3 (BioLegend; 1:100) and viability dye, in RPMI + 1% FBS for 30 minutes at 4'C and GFP+, GFP-CD45+CD3e+, GFP-CD45+CD3e- cells were sorted using BD FACSAria. Cells were then re-pooled at a combination of 50% CD45+CD3e+, 25% CD45+CD3e-, 25% GFP+CD45-CD3e-. Droplet-based 3' end single cell RNA sequencing was performed using the Chromium Controller and Illumina HiSeq 4000. Libraries were prepared using Chromium Single Cell 3' Reagent Kits v2 according to manufacturer's protocol (10X genomics). Generated libraries were sequenced on Illumina HiSeq 4000.
GEO
GSM4399494