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. Author manuscript; available in PMC: 2021 Nov 10.
Published in final edited form as: Cell Rep. 2021 Oct 26;37(4):109896. doi: 10.1016/j.celrep.2021.109896

Figure 5. The evolution of two putative Abd-B binding sites is necessary for the expression of bond in the EB.

Figure 5.

(A) An EB RNAi screen identifies Abd-B as a possible regulator of bond in the EB.

(B) RNAi knockdown of bond in the EB led to a big decrease in GFP expression driven by the D. melanogaster bc3 construct. Scale bar, 100 μm.

(C) JASPAR analysis identified three putative Abd-B binding sites in the D. melanogaster bc3 construct.

(D) Site-directed mutagenesis of these putative Abd-B binding sites individually shows that the Abd-B_1 and the Abd-B_2 sites are necessary for GFP expression driven by the bc3 construct, but not the Abd-B_3 site. Scale bar, 100 μm.

(E) Evolutionary analysis of the Abd-B_1 and the Abd-B_2 sites shows that all species have putative Abd-B binding sequences at Abd-B_1, and most species have putative Abd-B binding sequences at Abd-B_2 except D. virilis.

(F) Swapping in the D. virilis sequence at both sites either individually or in combination led to the loss of GFP expression in the hb of the EB driven by the D. melanogaster bc3i construct.