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. 2021 Apr 26;17(4):e1009536. doi: 10.1371/journal.pgen.1009536

Fig 1. Effects on expression due to eve insulator removal and replacement.

Fig 1

(A) Map of the eve pseudo-locus construct (inserted in either orientation, H5 or N5). The wild-type eve and TER94 coding regions were replaced with lacZ and EGFP, respectively. Rectangles represent regulatory elements: red, nhomie and homie insulators; yellow, Polycomb response elements (PREs); blue, embryonic stripe enhancers labeled with either numbers for early stripes, or “late” for the 7 later stripes; pink, tissue specific enhancers for: APR, anal plate ring; meso, mesoderm (other known elements are either unlabeled or not included). 5’P and 3’P: 5’ and 3’ P-element ends, part of the attP site previously inserted at cytological location 74A2 or 23C4. Note that the 5’P end contains a P-element promoter, but the 3’P end does not. (B) Inserted in the H5 orientation at site 74A2. Rows: lacZ expression (visualized by RNA in situ hybridization) at embryonic stages 5, 7, 11, 13. Columns: wt: “wild-type” intact pseudo-locus construct; Δhomie: 500 bp homie was replaced with a similar length of phage λ DNA; Δnhomie: 200 bp core nhomie was deleted. Affected tissues are labeled, including stripes number 1, 4, 5, and 6 at stg5, and mesoderm (meso) and anal plate ring (APR) at stage (stg) 11 and stg13. (C) Same as in A, but in the N5 orientation. Early stripe 3 at stg5 is marked (note the reduction in intensity of stripe 3 relative to flanking stripes in Δnhomie vs. the same comparison in wt). The 7 late stripes at stg7 are marked with dots (note the reduction in intensity in Δnhomie relative to wt). (D) In the H5 orientation at site 23C4. wt (top row) and Δhomie (bottom row) are shown at stg5 and stg11. The affected expression is marked with numbers and "meso". (E) In the N5 orientation at site 23C4. wt (top row) and Δnhomie (bottom row) are shown at stg5 and stg7. Affected expression is marked with “3” and dots. (F) 500 bp homie was replaced with either a Fab7, Fab8, or gypsy insulator in the H5 orientation. lacZ expression is shown at stages 5, 7, 11, and 13.