GSM5964422: APOB-specific Tet+ cells in individuals with and without sCVD, TCR; Homo sapiens; OTHER
Frozen PBMC samples were thawed and stained with APOB-p18 DRB1*07:01 tetramer after Fc blocking and dasatinib. After live dead aqua (1:1000, Thermo Fisher Scientific) staining (in PBS, 4℃ for 20min), cells were stained with surface marker antibodies on ice in the dark for 30min. Cells were washed twice with 2 mL flow buffer (PBS with 2%FBS, Gemini Bio). The cells were sorted by the FACS sorter (FACSAria, BD) into 4 Eppendorf tubes (DNA low bind), each of which contained 100μl flow buffer. Some surplus of the sorted Th1 and Tmem were discarded. Each single cell suspension in a 1.5mL sterile Eppendorf tube containing no more than 1 million cells in 50 μL of staining buffer (2% FBS in 1X PBS) was incubated with 2.0 μL of unique hashtag oligonucleotide antibody. Library was prepared per 10x Genomics protocol.