Figure 3.

Autoquin increases lysosomal mass and sequesters Fe²⁺ to the lysosomes in MCF7 cells, causing an increase in lysosomal reactive oxygen species. a) Fluorescence microscopy images showing the subcellular localisation of Fe²⁺ (red) and lysosomes (green) by means of RhoNox‐M and Lysotracker deep red (DR) fluorescence, respectively. b) Flow cytometry quantification of Lysotracker DR intensity from cells treated according to (a). c) Flow cytometry quantification of RhoNox‐M intensity from cells treated according to (a). d) Flow cytometry quantification of intracellular Fe²⁺ using an alternative turn‐on fluorescent probe RhoNox‐1. e) Fluorescence microscopy images showing the subcellular localization of ROS (red) by means of fluorogenic reaction with CellROX deep red in MCF7 cells treated with autoquin (5 μm) for 24 h, scale bar=10 μm. Colocalization with Lysotracker DND‐26 (green) assessed by Pearson correlation coefficient (R). ****p<0.0001, unpaired Student's t‐test.