Day 1: Drosophila cell transfection

Drosophila S2R+ were transfected with DNA (UAS-Luc, G4-DBD-Yki and pIZ-Rluc) using Effectene transfection reagent. The cells were then bathed with the dsRNA library (Ambion) and cultured at 25 °C, 0% CO₂ for 4 days.

Day 5: Cell lysis and dual-luciferase reporter assay

The culture media was aspirated and Passive Lysis Buffer was added.

Firefly luciferse and Renilla luciferase activity were quantified on the Envision plate reader.