Fig. 5 | Nature Communications

Fig. 5

From: Partial proteasomal degradation of Lola triggers the male-to-female switch of a dimorphic courtship circuit

Fig. 5The alt text for this image may have been generated using AI.

Lola29M/F molecular actions. a–h robo1 promoter-luciferase reporter assays. a The robo1 genomic map (upper) and reporter schematics (lower). The genomic distance (in kb) is indicated on the top with the reference point 0, at which the robo1 genomic fragment was fused with the luciferase-coding sequence (blue box with an arrow). Below the thick line, the exon (box)-intron (thin line) organization of robo1 is shown. Filled and open portions of the box represent the coding and non-coding regions, respectively. b The relative activities (ordinate) of a 4.1 kb robo1-promoter reporter cotransfected without (-) or with (+) Lola29M, FruBM or both. c The activities of reporters (ordinate) carrying a robo1-promoter fragment of different lengths (the fragment length is indicated on the abscissa) with (filled bars) or without (open bars) cotransfection of Lola29M or truncation-resistant Lola29M[K41R]. d The reporter activities were unaffected by cotransfection with Lola29F-like (Lola29M[Δ1-300]). e The suppression effect of Lola29M on reporter activities was inhibited by Lola29M[Δ1-300]. f–m Lola29M binding to the robo1 promoter. f Genomic fragments used for EMSA. Probe DNA B and Probe DNA B ∆DR1 are shown in an expanded scale in the second and third rows. Reporters are schematically illustrated at the bottom. Pal 1: palindrome sequence, FROS: the FruBM-binding region, DR1: direct repeat 1. g The sequence around DR1 and FROS. The deletion in the 0.9 kb reporter is also indicated. h The repressor action of Lola29M[K41R] on the 0.9 kb reporter with or without DR1. *P < 0.05 by the Mann–Whitney U test. i–m EMSA with fragment-B detected a retarded band (arrowhead) in the presence of truncation-resistant Lola29M[K41R] (lane 2), which was eliminated by adding the unlabeled fragment-B as competitors (lane 3). Mouse IgG had no effect (lane 5), whereas the Lola29M[K41R]-recognizing anti-V5 induced a super-shift (asterisk; lane 6). The ∆DR1 probe did not induce retardation (lanes 8). An arrow indicates free DNA. Lola29M failed to produce the retarded band in the presence of Lola29F (lane 11), which on its own, did not produce the retarded band either (lane 13). Source data are provided as a Source Data file

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