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Fig. 2. bHLH repressor-encoding genes of the E(spl)-C are also subject to direct transcriptional repression by Su(H) in SOPs. (A) Diagrams of rescue fragment and enhancer-reporter constructs for E(spl)m8. (1) Genomic DNA fragment including the E(spl)m8 gene, with protein coding sequence (blue) and positions of Su(H) (S) and proneural protein (E) binding sites marked. Arrow indicates transcription start site. (2,3) E(spl)m8 enhancer-GFP reporter constructs with Su(H)-binding sites intact (2) or mutated (Sm; mutant sites indicated by a red cross) (3). Minimal Hsp70 promoter is shown. (B-G) Wing discs from late third-instar larvae carrying one copy of either E(spl)m8-GFP (B-D) or E(spl)m8 Sm-GFP (E-G). (B,D,E,G) GFP expressed from E(spl)m8 enhancer-reporter transgenes (green). (C,D,F,G) Anti-Hnt antibody staining (magenta) marks SOPs. Insets in merged D and G show higher-magnification views of the boxed regions of the two wing discs. (B) E(spl)m8-GFP is strongly expressed in PNCs, but activity is restricted to non-SOPs (D; inset: SOPs in top and bottom clusters are obstructed by non-SOPs). (E) Mutation of Su(H)-binding sites causes severe or complete loss of reporter expression in non-SOPs, but strong ectopic expression in SOPs (G).

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