| Screen Status: | Primary |
| Screen Overview: | Dual Luciferase-Renilla Genome-wide screen using 2 replicates in 384-well plates. |
| Screener: | Filipe Josue (Tapon Lab). Extension:3661, Rm 230 |
| Library: | Ambion Genome Library |
| Library Description: | Drosophila genome library in 36 384-well plates from Ambion |
| Library Supplier: | Ambion |
| Library Format: | 36 plate(s) in 384 well format |
| Replicate Info: | 2 replicates |
| Publications: | Combined functional genomic and proteomic approaches identify a PP2A complex as a negative regulator of Hippo signaling.. Ribeiro, Josue et al., 2010 |
| Keywords: | Hippo signaling |
| Cell Line: | S2R+ |
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.