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