Screen Status: Primary
Screen Overview: Primary Screen
Screener: Franzi Baenke (Schulze Lab).
Extension:2049, Rm 113
Library: Glucose Metabolism Custom Franzi
Library Description: Re-arrangement of Glucose Metabolism library (libID 95) with additional siRNAs ordered seperately. 3 plates
Library Supplier: Dharmacon
Library Format: 3 plate(s) in 96 well format
Replicate Info: 3 replicates
Publications: Functional screening identifies MCT4 as a key regulator of breast cancer cell metabolism and survival.. Baenke et al., 2015
Keywords: Breast Cancer, Metabolism, Viability
Cell Line: MCF-7

Screen Method

Day 1: Transfect cell lines

15 breast cancer cell lines and 3 normal breast cell lines are transfected using Lullaby and custom designed metabolic enzyme library for 1 day under normal and hypoxia conditions.

Day 2: Change the media

Change for fresh culture media and culture the plates for further 3 days under the respective normal and hypoxia condition.

Day 5: Fix the plates with 80% EthOH and stain

Stain with capase-3 antibody to detect the Apotosis and stain with DAPI for nuclear counting.

Readout

Acumen Explorer eX3 laser scanning cytometer was used to determine cell number per well and Caspase-3 intensity.