Extended Data Fig. 2: Schematic summary of sorting and hashtag oligo (HTO) staining. | Nature Cardiovascular Research

Extended Data Fig. 2: Schematic summary of sorting and hashtag oligo (HTO) staining.

From: Single cell transcriptomics and TCR reconstruction reveal CD4 T cell response to MHC-II-restricted APOB epitope in human cardiovascular disease

Extended Data Fig. 2

Tet+ cells, Th1 cells, Treg, and CXCR3-memory T cells (Tmem) were sorted into 4 different tubes, and were stained with antibodies with hash tag oligo (HTO1-4). Treg, Th1, and Tmem were pre-gated for CD45RA-. After staining of HTO antibody, tet+ cells weren’t washed, not to lose any cells, because one wash would lose almost half of the cells. Other cells (Th1, Treg and Tmem) were washed three times following to the manufacturer’s instruction. The volume of each HTO antibody had previously been titrated. The cell number of these three cell types (Th1, Treg and Tmem) was appropriately adjusted, and they are merged into the tet+ cell tube. At the same time, bulk CD4 T cells from a healthy donor were merged into the tube for the following batch correction. After that, the sample proceeded to barcording, cDNA amplication, library preparation, and sequencing.

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