GSM8995481: D7_sc; Mus musculus; RNA-Seq
Treg cells were multiplexed together from SLO or tumor. For more details, refer to Methods. RNA sequencing was performed using 10X genomics. For tumor: cells were hybridized for 18 hours, washed, and encapsulated in one lane targeting 60,000 cells. Indexing PCR steps were performed using 9 amplification cycles for both the gene expression and cell surface protein libraries. Final sequencing libraries were sequenced on an Illumina NovaSeq X Plus System (index 1: 10 cycles, index 2: 10 cycles, read 1: 28 cycles, read 2: 88 cycles). For SLO: Approximately 30,000 cells were targeted. Final libraries were sequenced on Illumina NovaSeq 6000 - S4 platform (R1 – 28 cycles, i7 – 8 cycles, R2 – 90 cycles). Viable cells were identified on the basis of library size and complexity, whereas cells with >20% of transcripts derived from mitochondria were excluded from further analysis.