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.