Fig. 3: Combining mini-yellow rescue and sex conversion.

A Structure of the YRN (yellow rescue with nix) construct created by adding nix 3&4 isoform, flanked by its native regulatory region and an SV40 poly(A) terminator motif (red), to the HspYR (Heatshock yellow rescue) plasmid, which contains the hsp83-driven mini-yellow and the eGFP transformation marker. B–D Crossing scheme for isolating functional integrations. B Transgenic male isofounders are crossed with yellow females. C F1 progeny were analyzed for sex and transgenic status. D F2 transgenic males from isofamilies with significant sex bias (χ² ≥ 15.1, p < 0.0001; indicated by *) were individually crossed to yellow females to exclude subfamilies producing transgenic females or yellow individuals and to purify M-locus free male lineages by molecular genotyping for the endogenous nix and yellow. E Sex conversion and yellow rescue in crosses between YRN and yellow females, using HspYR as a control. The frequency of pigmentation phenotypes (dark and yellow), separated by sex, is presented as the mean proportion ± SEM across three gonotrophic cycles from two cages. Numbers indicate the total individuals recorded per group. Source data are provided in the Source Data file.