| Screen Status: |
Primary |
| Screen Overview: |
Genome screen using 384 well library |
| Screener: |
Marco Gerlinger (Swanton Lab).
Extension:3462, Rm 115 |
| Library: |
Full Genome 384 |
| Library Description: |
Full Dharmacon 267 plate genome library stamped into 384 well plates |
| Library Supplier: |
Dharmacon |
| Library Format: |
67 plate(s) in 384 well format |
| Replicate Info: |
3 replicates
Biological replicates |
| Publications: |
Genome-wide RNA interference analysis of renal carcinoma survival regulators identifies MCT4 as a Warburg effect metabolic target.. Gerlinger et al., 2012
|
| Keywords: |
Everolimus, Kidney cancer, Viability |
| Cell Line: |
RCC4 |
Day 1: Transfection of RCC4, A498 and 786-O cell lines
Cells were transfected with genome siRNA library (Dharmacon) using Dharmafect3 (RCC4) or LipoRNAiMAXi (A498 and 786-O) and cultured for 1 day.
Day 2: Addition of Everolimus
Everolimus is added on top of the culture media. For control plates, fresh media only was added. The plates were then cultured for further 3 days.
Day 5 Plates are fixed with 80% EthOH and stained with DAPI
Plates were fixed with 80% Ethanol and stained with DAPI.
Readout
Acumen Explorer eX3 laser scanning cytometer was used to determine cell number per well.
