Research Paper Volume 16, Issue 2 pp 1925—1937

Melatonin improves stroke through MDM2-mediated ubiquitination of ACSL4

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Figure 3. MDM2 affects ferroptosis by regulating ACSL4 protein levels, not mRNA levels. (A) Ubibrowser database is used to predict the ubiquitin E3 ligases that ACSL4 may interact with. (B) QRT-PCR shows overexpression and knockdown efficiency of MDM2 in HT-22 cells. (C) CCK-8 assay shows MDM2 overexpression increased while MDM2 knockdown decreased the IC50 of RSL3 in HT-22 cells. (D) DCFH-DA staining shows MDM2 overexpression inhibited while MDM2 knockdown increased ROS levels in HT-22 cells treated with RSL3. (E) QRT-PCR shows MDM2 overexpression or knockdown did not affect ACSL4 mRNA levels. (F) Western blot shows MDM2 overexpression decreased while MDM2 knockdown increased ACSL4 protein levels. Data are presented as mean ± SD. *P < 0.05 vs. NC or si-NC group.