Fig. 6. Pharmacologic depleting of the free iron reservoir attenuates the DHA/DDP-induced ferroptosis.

A PANC1 cells were treated with DHA/DDP (45 μM) in the presence or absence of DFO. B PANC1 cells were treated with DHA/DDP (30 μM) in the presence or absence of DFO and Fe²⁺, cell survival was detected by CCK8. C, D After indicated treatment, PANC1 cells were loaded with RPA or DCF-DA probe for 30 min, and fluorescence intensities were detected by microplate spectrophotometer and normalized to the corresponding cell number. E MitoTracker Red labeled pancreatic cancer cells were subjected to the confocal microscope for observing the changes of mitochondrial morphology after the indicated treatment. Scale bars: 10 μm. F, G OCR of PANC1 cells was carried out with a Seahorse analyzer after the addition of oligomycin, FCCP, and Antimycin A/Rotenone. The basal respiration and ATP production were calculated on the right. H, I To assess lipid ROS production, pancreatic cancer cells were treated with DHA and DDP with or without DFO, fer-1. Treated cells were loaded with BODIPY C11 probe for 30 min followed by confocal laser microscope. Scale bars: 50 μm. Statistical results of the fluorescence intensities were shown on the right (values represented mean ± SD. ^★P < 0.05, ^★★P < 0.01 versus control).