Journal: bioRxiv

Article Title: A recombinant dilp2GS-rpr donor line for adult-inducible IPC ablation across Drosophila genetic backgrounds

doi: 10.64898/2026.06.17.733056

Figure Lengend Snippet: (A) Crossing scheme used to generate the recombinant donor chromosome. The UAS -rpr responder line was crossed to the dilp2 GS -GAL4/CyO driver line. F1 females carrying dilp2 GS -GAL4 and UAS -rpr on homologous second chromosomes were crossed to sna[Sco] / CyO males to recover recombinant chromosomes generated during female meiosis. Candidate F2 flies carrying a recombinant second chromosome over CyO were selected and the resulting donor chromosome was backcrossed to the matched-control background for seven generations. (B) Representative F2 flies from the recombinant-screening cross. The orange-eyed fly was selected as a candidate dilp2 GS-rpr recombinant carrier, whereas a lighter/yellow-eyed F2 sibling is shown for comparison. Eye color was used as a screening marker for candidate recombinant flies. (C) Schematic of RU486-inducible IPC ablation in the dilp2 GS -rpr donor line. The dilp2 promoter drives GeneSwitch-GAL4 expression in adult insulin-producing cells. In the absence of RU486, GeneSwitch-GAL4 is expected to remain inactive and UAS-rpr is not induced. After RU486 treatment, activated GeneSwitch-GAL4 drives rpr expression from the UAS responder, leading to Reaper-mediated IPC ablation. (D) PCR genotyping of parental and candidate donor lines. Lane 0, no-template genomic DNA control; lane 1, dilp2 GS - GAL4 parental line; lane 2, UAS -rpr parental line; lane 3, candidate dilp2 GS -rpr recombinant donor line. Expected PCR products are 328 bp for dilp2 GS - GAL4 and 805 bp for UAS -rpr . Detection of both products in the candidate donor line supports the presence of both transgenic elements; amplified products were further verified by Sanger sequencing.

Article Snippet: RU486 (mifepristone; 98% purity; Thermo Scientific Chemicals, TS45998-0050) was dissolved in 80% ethanol and added to food at a final concentration of 200 μM.

Techniques: Recombinant, Generated, Control, Comparison, Marker, Expressing, Transgenic Assay, Amplification, Sequencing

Journal: bioRxiv

Article Title: A recombinant dilp2GS-rpr donor line for adult-inducible IPC ablation across Drosophila genetic backgrounds

doi: 10.64898/2026.06.17.733056

Figure Lengend Snippet: (A–D) Whole-body glucose levels in female recombinant dilp2 GS -rpr line and the background-matched control line after 8, 10, 12, or 14 days of RU486 or vehicle treatment. (E) Glucose levels across the 8-, 10-, 12-, and 14-day treatment time course in the recombinant dilp2 GS -rpr line and the background-matched control line treated with RU486 or vehicle. (F, G) Short-term dye-based food intake measured by blue dye feeding assay in male (F) and female (G) recombinant dilp2 GS -rpr line and the background-matched control line treated with RU486 or vehicle. (H) Relative dilp2 mRNA expression measured by qPCR in adult female recombinant dilp2 GS -rpr flies treated with RU486 or vehicle. Expression was normalized to Actin5C and calculated using the 2 -ΔΔCt method. (I–K) Whole-body glucose (I), glycogen (J), and triglyceride (K) levels in female flies from the recombinant dilp2 GS -rpr line and the background-matched control line after 8 days of RU486 or vehicle treatment. Data are presented as mean ± SEM. RU+, RU486-treated; RU−, vehicle-treated. Statistical comparisons shown above bars compare RU486-treated flies and the corresponding vehicle-treated controls within each line. * P < 0.05, ** P < 0.01, *** P < 0.001; n.s., not significant.

Article Snippet: RU486 (mifepristone; 98% purity; Thermo Scientific Chemicals, TS45998-0050) was dissolved in 80% ethanol and added to food at a final concentration of 200 μM.

Techniques: Recombinant, Control, Feeding Assay, Expressing