Fig 2: Expression of selected proteins of interest in primary trophoblast cells. Western blot analysis was carried out to evaluate the protein expression of cholesterol efflux transporters ABCA1 (A) and ABCG1 (B), glucose uptake transporter SLC2A1 (C), 4F2 heavy chain SLC3A2 (D) and amino acid transporter SLC7A5 (E), and iron uptake receptor TfR1 (F). Protein expression was normalized to the total protein, as visualized by Ponceau S staining. Data are shown as median with IQR; n = 4 for each stage. Asterisks indicate significance according to nonparametric t-test (Mann-Whitney test): *p ≤ 0.05, **p ≤ 0.01.

Fig 3: Iron accumulation in mice containing the S47 variant.a Domain model of TP53 showing the positions of S47 P47 and R72 SNPs studied in this paper. b Iron levels in P47 and S47 human lymphoblastoid cell lines and mouse embryonic fibroblasts measured by loss of calcein fluorescence. Mean fluorescence intensity reflects iron levels in untreated (black) or 100 µm DFO-treated (gray) MEFs and LCLs (n = 3 biological replicates). Error bars represent means ± s.e.m. **P < 0.01, **P < 0.01, *P < 0.05, by unpaired Student’s t test, relative to P47 mice. c Formalin-fixed sections of male S47 and P47 mouse spleens stained by Prussian Blue; representative images are of six biological replicates per group with 5–7 fields per sample. d Percent viability of P47 (white) and S47 (black) monocyte-derived macrophages, 48 h after treatment with ferroptosis inducers Erastin and RSL3. (n = 3 biological replicates each with three technical replicates). Error bars represent means ± s.e.m. ***P < 0.001, **P < 0.01, *P < 0.05, ns—not significant; by unpaired Student’s t test. e Relative levels of the iron transport protein Tfrc in P47 and S47 mouse MDMs measured by immunoblot with or without DFO treatment (representative blot of three technical replicates). f ICP-MS shows significantly higher iron content in S47 male mouse splenocytes (n = 10 biological replicates). Error bars represent means ± s.e.m. ***P < 0.001, **P < 0.01, *P < 0.05, ns—not significant; by unpaired Student’s t test, relative to P47 mice. g CBC profiles in S47 and P47 male mice (n = 8 biological replicates). Error bars represent means ± s.e.m. **P < 0.01, *P < 0.05, by unpaired Student’s t test, relative to P47 mice. h Allelic frequencies for the S47 SNP in populations of African descent; data are obtained from the 1000 Genomes Project (left panel). Associations between TP53 rs1800371 (S47) in individuals with high serum ferritin level and/or high saturated transferrin receptor in African Americans from the HEIRS database (right panel). (MAF = minor allele frequency, OR (95% CI) = odds ratio (95% confidence interval), * from25. Source data are provided as a Source Data file.

Fig 4: The alterations in iron metabolism, Nrf2/HO-1 signaling, and EMT process in the kidneys of CKD mice, which could be attenuated by AST-120. (A) Western blot analysis and quantification for iron metabolism-related Tf, TfR, FTH, DMT1, FPN, and NCOA4 protein expression in the kidneys. (B) The IHC staining and quantification for FTH in the kidneys; scale bar = 50 μm. (C) Western blot analysis and quantification for intracellular antioxidant system-related Keap-1, Nrf2, and HO-1protein expression in the kidneys. (D) The IHC staining and quantification for HO-1 in the kidneys; scale bar = 50 μm. (E) Western blot analysis and quantification for EMT-related fibronectin, E-cadherin, and vimentin protein expression in the kidneys. Data are presented as median (minima and maxima) (n = 6 mice/group). *, p < 0.05, compared with the control (C) group. #, p < 0.05, compared with the adenine (Ade) group. The Kruskal–Wallis test was used to analyze statistical differences.