About 400 0-14 hour old embryos per biological replicate (three biological replicates prepared per genotype) were dechorionated, dounced in 5 ml crosslinking solution (50 mM Hepes pH 7.9, 1 mM EDTA, 0.5 mM EGTA, 100 mM NaCl, 1.8% formaldehyde), and fixed for 15 minutes. Nuclei were sonicated in RIPA buffer in a Covaris S220 sonicator as described in Kaushal et al. 2021. 60 third instar larval central nervous systems per biological replicate (two biological replicates per genotype) were dissected in PBS and chromatin was prepared as in Kaushal et al. 2021. NEBNext Ultra II DNA Library Prep kit for Illumina