酚红 (PR) 是培养基中常用的 pH 指示剂化合物。然而，尚不清楚该化合物是否可以干扰铁死亡的药理诱导。在这里，使用高内涵活细胞成像死亡分析，我们确定培养基中 PR 的存在使正常细胞和肿瘤细胞预先适应咪唑酮erastin (IKE) 或 GPX4 阻断介导的系统 xc 抑制诱导的铁死亡。与 RSL-3 相比，但对内过氧化物 FINO2 治疗没有显着影响。从机制上讲，我们揭示了 PR 降低了抗铁死亡基因 Slc7a11、Slc3a2 和 Gpx4 的水平，同时促进了铁死亡关键诱导物 de Acls4 的过度表达。此外，通过超分辨率分析，我们确定 PR 的存在使系统 xc- 与质膜错位。因此，我们的结果表明，培养基中 PR 的存在对于准确解释细胞对 IKE 或 RSL-3 介导的铁死亡诱导的敏感性可能是一个有问题的伪影。版权所有 © 2024。由 Elsevier Inc. 出版。

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 xc- 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.Copyright © 2024. Published by Elsevier Inc.