GSM9150434: KP cells, WT, replicate 1; Mus musculus; OTHER
CUT&Tag experiment was conducted using CUT&Tag kit complemented with  Hyperactive TF Enhancer Module  from Vazyme (TD904-C2). Briefly, cells were collected and treated with TF enhancer. Subsequently, cells were bound with Concanavalin A-coated magnetic beads, and the plasma membrane was permeabilized with digitonin. Through the use of a primary antibody targeting HA or non-targeting IgG control, a corresponding secondary antibody, and Protein A/G, the transposase fused to Protein A/G was guided to precisely cleave DNA sequences near the target protein. During cleavage, adaptor sequences were added to both ends of the DNA fragments, which were then amplified by PCR to generate libraries for high-throughput sequencing. CUT&Tag libraries were prepared using the CUT&Tag kit complemented with  Hyperactive TF Enhancer Module  (TD904-C2) and TruePrep Index Kit V2 for Illumina (TD202) from Vazyme. All of the cleaved DNA extracted using CUT&Tag kit was used as starting material for anti-HA and anti-IgG samples. Before PCR anplification, 0.05 pg DNA Spike-in was added into each sample as a reference standard, primarily for inter-sample normalization of the sequencing data. Libraries were then amplified using 16 cycles on the thermocycler. Libraries ere validated using the Fragment Analyzer 5300 system.