Fig. 1.

Inhibition of the mevalonate pathway triggers oxidative stress and ferroptosis in PDA. A, KP4 cells were treated with simvastatin (20 μM) w/or w/o supplementation with mevalonate (500 μM) for 2 h and stained for the detection of cytoplasmic (DCFDA, green) or mitochondrial (MitoSOX, red) ROS. Nuclei counterstained with HOECHST. Representative images (quantification in Suppl. Fig. 1). B–C, Flow cytometry analysis of KP4 treated with simvastatin (20 μM) showing enhanced ROS (B) and lipid peroxides (C) levels. D, KP4 cells were treated with simvastatin (20 μM) w/or w/o supplementation with mevalonate (500 μM) and incubated with C11-BODIPY after 1 h. Representative images of split channels (red: reduced C11; green: oxidized C11) and merge. E-F, KP4 cells were treated with simvastatin (20 μM) and/or Ferrostatin-1 (2 μM) and stained with C11-BODIPY (E). Graph shows the ratio between oxidized and reduced C11, quantified by flow cytometry (ratio of mean fluorescence intensity (MFI) for each sample). In F, cells were counted after 2 days. For all panels, experiments are representative of 2 independent biological repeats. Dots denote biological replicates. Bars show mean, ±SD (*, P < 0.05; **, P < 0.01; ***, P < 0.001; calculated over vehicle-treated cells unless otherwise indicated). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)