Live Cell Imaging of Spatiotemporal Ca²⁺ Fluctuation Responses to Anticancer Drugs

Synthetic triterpenoids, 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) and its derivatives are known to have potent anti-cancer and anti-inflammatory activities. The mechanisms of actions of CDDO and its derivatives as potential therapeutics remain elusive. Previous studies found that CDDO-Me triggers apoptosis by inducing extracellular Ca²⁺ influx followed by endoplasmic reticulum (ER)-derived vacuolation. Since Ca²⁺ activity in cells is dynamic and needs to be tracked in real time in living cells, we report a high-throughput and high-content imaging method to track CDDO-induced Ca²⁺ fluctuation in both ER and cytosol with MATLAB script for data analysis and visualization.