(A) Schematic of midgut lineage progression indicating cell types examined in this study. (B) Chromatin accessibility displays expected trends at the escargot locus, known to be expressed exclusively in ISCs and EBs, but not ECs. Upstream promoter region shows greatest chromatin accessibility in ISCs, compared to other cell types. Similarly, dynamic peaks are observed in both 3’ and 5’ distal regions (putative enhancer regions), which are absent in ECs. y-axes = reads per million (rpm). (C) Chromatin accessibility at the nubbin locus, known to be expressed exclusively in ECs. y-axes = reads per million (rpm). (D) Hierarchical clustering of differentially accessible regions in gut cell types. Major clusters are observed in which accessible chromatin is enriched specifically in either ISCs or ECs, whilst smaller clusters indicate fewer regions with up or down-regulated accessibility in EBs. (E) Principal component analysis (mean of all replicates) indicates distinct groupings of both lineages. (F) Correlation matrix (Spearman’s rank) of means of all cells in CNS and midgut lineages. Individual lineages denoted with red outline. Note relatively high correlation between NSC and ISC (Asterisk – R2 = 0.76), whilst NSC correlation with EC and adult neurons are comparable.