Motif-enrichment analysis identified binding sites for XBP1s in Neo-2/15 stimulated BBζ NK cells following co-cultured with AsPC-1 cells

Motif-enrichment analysis identified binding sites for XBP1s in Neo-2/15 stimulated BBζ NK cells following co-cultured with AsPC-1 cells, as determined using CUT&TAG-seq, were mainly at the gene encoding nuclear respiratory factor 1 (NRF1) The CUT&Tag assay was conducted using Hyperactive Universal CUT&Tag Assay Kit for Illumina. Briefly, 106 BBζ NK cells were collected after co-cultured with AsPC-1 cells and washed. BBζ NK cells were bound to ConA beads for 10 min at 25 °C. Then cells were incubated with H3K9me2 antibody at 4 °C overnight. Anti-mouse IgG was added and incubated for 1 hour at 25 °C. Then cells were washed and incubated with 0.04 μM pA/G–Tnp for 1 hour at 25 °C and washed three times with DIG 300 buffer, resuspended in tag mentation buffer and incubated at 37 °C for 1 hour. Tag mentation was stopped by adding proteinase K, buffer LB and DNA extract beads. After incubation at 55 °C for 10 min, cells were plated on a magnet and unbound liquid was removed. Beads were rinsed twice. Libraries were constructed using TD903 Hyperactive Universal CUT&Tag Assay Kit for Illumina, and TD202 TruePrep Index Kit V2 for Illumina (Vazyme Biotech).