1. The phenol red compound: A potential artifact in pharmacological induction of ferroptosis.
- Author
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Vera M, Barahona MJ, Nova-Lamperti E, Nualart F, and Ferrada L
- Subjects
- Humans, Piperazines pharmacology, Amino Acid Transport System y+ metabolism, Amino Acid Transport System y+ genetics, Artifacts, Imidazoles pharmacology, Cell Line, Tumor, Culture Media chemistry, Animals, Carbolines, Ferroptosis drug effects, Ferroptosis genetics, Phospholipid Hydroperoxide Glutathione Peroxidase metabolism, Phospholipid Hydroperoxide Glutathione Peroxidase genetics, Phenolsulfonphthalein metabolism
- Abstract
Phenol red (PR) is a commonly used compound in culture media as a pH indicator. However, it is unknown whether this compound can interfere with the pharmacological induction of ferroptosis. Here, using high-content live-cell imaging death analysis, we determined that the presence of PR in the culture medium preconditioned normal and tumor cells to ferroptosis induced by system x
c - inhibition mediated by imidazole ketone erastin (IKE) or GPX4 blockade in response to RSL-3, but had no significant effects against treatment with the endoperoxide FINO2 . Mechanistically, we revealed that PR decreases the levels of the antiferroptotic genes Slc7a11, Slc3a2, and Gpx4, while promoting the overexpression de Acls4, a key inducer of ferroptosis. Additionally, through superresolution analysis, we determined that the presence of PR mislocalizes the system xc - from the plasma membrane. Thus, our results show that the presence of PR in the culture medium can be a problematic artifact for the accurate interpretation of cell sensitivity to IKE or RSL-3-mediated ferroptosis induction., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)- Published
- 2024
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