GSM8463926: M25+ Pseudobulk; Mus musculus; RNA-Seq
Tetramer+ cells were sorted into 1.5 mL collection tubes containing 500 μl of PBS:FBS (1:1) supplemented with RNase inhibitor (Takara, Mountain View, Ca, USA; 1:100). After sorting, ice-cold PBS was added to reach a final volume of 1400 μl. The cells were centrifuged (400 x g, 5 minutes, 4°C), cell suspension were incubated with the 10x Genomics Master Mix, loaded onto a Chromium Next GEM Chip (10x Genomics, Pleasanton, CA, USA) Library construction was performed according to the Chromium Next GEM Single Cell 5' Protocol. Samples were processed using 10X v2 chemistry as per the manufacturer's recommendations; 11 and 12 cycles were used for cDNA amplification and library preparation, respectively.