show Abstracthide AbstractWe performed ATAC-seq using 2-day old female and male flies for head and testes samples, respectively, from D. melanogaster (Bloomington #2057), D. simulans (w501), and D. yakuba (Tai18E2), as well as head ATAC-seq of D. suzukii. For each experiment, 25 newly emerged flies per replicate were collected and transferred to new vials. There were three replicates per sample. Two days later, we dissected heads or testes in cold PBS. Sample preparation was performed as described (Witt et al., 2021). Libraries were constructed using the same primers as (Buenrostro et al., 2015) and sequenced on a 75-bp paired-end Hiseq X platform.