GSM4678095: pMIG CAR TILs_rep1 ATACseq; Mus musculus; ATAC-seq
50,000 GFP+CD8+Thy1.1+ CAR T cells  were sorted from B16-huCD19 tumors or or 50,000 CD8+ T cells were sorted from in vitro cultured CD8 T cells. ATAC-seq libraries were prepared following the omni-ATAC protocol with minor modification6. 50,000 cells were collected by sorting and washed twice with cold-PBS at 600g for 5 minutes. Cell pellets were resuspended in 50 µl ATAC-lysis buffer (10 mM 1 M Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% NP40, 0.1% Tween 20, 0.01% Digitonin ), and incubated on ice for 3 minutes, after which 1 ml washing buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1%Tween 20) was added, and the cells were spun down at 1000g for 10 minutes at 4℃ . The supernatant was removed carefully, and the nuclei were resuspended in 50 µl of transposition mix (25l of TD buffer [20 mM Tris-HCl pH 7.6, 10 mM MgCl2, 20% Dimethyl Formamide], 2.5 l of 2 µM transposase 16.5 l PBS, 0.5l 1% digitonin, 0.5l 10% Tween-20, 5l water) and incubated at 37℃ for 30 minutes. DNA was purified with a Qiagen MinElute Kit (Qiagen). ATAC-seq, 2x50- cycle paired-end sequencing (illumina)
GEO Accession
GSM4678095