title: A Het rep 2
design: CUT&Tag was performed on freshly harvested NPCs following the ConA-bead version of the Kaya-Okur and Henikoff (2020) protocol31, with minor modifications. Briefly, 200,000 freshly collected NPCs per sample were bound to ConA-coated magnetic beads. After washing in digitonin-containing buffers, cells were incubated with rabbit anti-POGZ (EPR10612) or rabbit IgG control antibodies overnight at 4C. Following secondary antibody binding, samples were treated with protein ATn5 transposase pre-loaded with Mosaic End adapters, incubated for one hour, and tagmented at 37C for one hour. The reaction was halted using EDTA, SDS, and ProteinaseK, followed by DNA purification with column cleanup. Libraries were amplified using 14 cycles of PCR with Q5 polymerase, purified via SPRI beads, and pooled for high-throughput sequencing on an Illumina NextSeq2000 (2150bp reads). sequencing was performed in biological triplicates to ensure reproducibility.