Single-cell RNA sequencing Analysis of Treg cells from mouse B16 melanoma tumors

Based on single-cell RNA sequencing technology, we studies gene expression profiles in Treg cells isolated from mouse tumors before and after rMVA therapy. We identified differential Treg populations from tumors before and after rMVA therapy. The difference of Treg clusters in tumors were also identified. Foxp3-GFP mice were intradermally implanted with B16 cells at 5x10^5 cells at the right side. After 7-10 days, tumors were dissected and digested into single cells. After staining with antibodies against CD45, CD3, CD4, GFP+ cells were sorted in BD FACSAria cell sorter and applied into Chromium Single Cell V(D)J Solution from 10x Genomics according to the manufacturer’s instructions. Cell-barcoded 5′ gene expression libraries were sequenced on an Illumina No-vaSeq6000 system.