GSM6569773: BMDM, PREP, binding, M1; Mus musculus; OTHER
NovoNGS® CUT&Tag 3.0 High-Sensitivity Kit (NovoProtein, Cat. #N259) was used for CUT&Tag library preparation according to the manufacturer's recommendations. Briefly, 1 × 105 cells were bound to Concanavalin A (ConA)–coated magnetic beads and were incubated with a primary rabbit anti-prolyl endopeptidase (1:100, Abcam, CAT. #ab58988) for 2 h at room temperature (RT). Then cells were shifted to incubation with ChiTag Goat anit-Rabbit IgG antibody (1:200) antibody for 1 h at RT. After washing by Wash buffer, cells were incubated with NovoNGS® ChiTag™ pAG-Tn5 for 1 h at RT. After washing by ChiTag Buffer, cells were resuspended in 50 μl of Tagmentation Buffer and incubated for 1 h at 37°C, followed by heat inactivation at 55°C for 10 min to stop the reaction. DNA fragments were extracted by Tagment DNA Extract Beads and amplified by using N5&N7 primers and 5×AmpliMix. After DNA purification by NovoNGS® DNA Clean Beads, libraries were sequenced on Illumina NovaSeq 6000 for generation of 150-bp paired-end reads.