FIP-gts and chloroquine cytotoxicity were determined by evaluating CCK-8 assay.  Using CCK-8 assay, chloroquine increased FIP-gts-induced cytotoxicity in parental and cisplatin-resistant urothelial cancer cell lines. 
CCK-8 assay
Cell death pathways, ROS and cell cycle arrested were analysed through flow cytometry and Western blot.  On flow cytometry, chloroquine enhanced FIP-gts-mediated sub-G1 accumulation, annexin V positive signal and mitochondrial membrane potential loss.  Using H2DCFDA staining and flow cytometry, FIP-gts and chloroquine did not induce ROS production. 
flow cytometry
Cell death pathways, ROS and cell cycle arrested were analysed through flow cytometry and Western blot. 
Western blot
ShRNA targeting to autophagy-related genes were tested to evaluate their autophagic cell death for resistant urothelial cells.


 To elucidate the role of autophagy in caspase-independent cell death by FIP-gts and chloroquine, LC3 shRNA were used to inhibit autophagy in N/P cells. 
ShRNA
Using H2DCFDA staining and flow cytometry, FIP-gts and chloroquine did not induce ROS production. 
H2DCFDA staining