HAMP Antibody - #DF6492

Fig. 7. Iron efflux blockade contributed to LPS/iE-DAP-induced ferroptotic cell death. (A) Gene expression of HAMP and (B–C) protein expression of DMT1, TFR, FPN, and hepcidin in hepatocytes treated with or without LPS/iE-DAP; n = 6 per group. (D) Cellular Fe2+ content. (E–F) Protein expression of FPN, hepcidin, ACSL4, ALOX15, and GPX4 in indicated groups; n = 3 per group. (G–I) Fluorescence images of lipid ROS observed by confocal microscope (Scale bar = 100 μm) and proportion of lipid ROS positive cells analyzed by flow cytometer; n = 3 per group. (J) Cell viability, (K) MDA content, and (L) GSH content in different groups; n = 4–6 per group. (M) Coimmunoprecipitation and ubiquitination analysis of FPN in hepatocytes; n = 3 per group. Data are presented as mean ± SEM.

Figure 3. 4-OI mitigates APAP-induced dysregulation of iron metabolism. (A) Pathways associated with genes/proteins interacting with differential metabolites were analyzed using the R package CePa. logP (x-axis): negative log of the enrichment p-value for drug bias-related metabolites. foldP (y-axis): ratio of enrichment p-values for efficacy-related vs. bias-related metabolites. The red circle highlights the ferroptosis pathway, which is identified as the most prominent among all analyzed pathways. (B) Hepatic iron content (n = 3). (C–I) Representative Western blot images and quantification of transferrin, TFR1, FTH1, FTL1, FPN1, and hepcidin protein levels in mouse liver tissues (n = 3). (J–L) A RT-qPCR assay determined the mRNA expression of FTH1, FTL1, and FPN1 (n = 6). (M) Representative immunohistochemical staining images showing FTH1, FTL1, and FPN1 expression. Scale bar: 100 μm, n = 3. GAPDH was used as internal loading control.

Figure 5 Iron levels in the hippocampus of CIH mice. (A) Perls’ staining in the hippocampus (Scale bar = 25 µm). (B) The mean density of Fe content as shown in panel A (n = 3). (C) The total iron content in the hippocampus tissue measured by ICP‒MS (n = 3). (D) The λ absorption of HuA with Fe2+ and Fe3+, respectively. (E) The expression of hepcidin mRNA (n = 5). (F) The sections were labelled for DAPI (blue), GFAP (green), and hepcidin (red) (scale bar = 25 μm, n = 3). (G) The expression of TfR1 proteins measured by Western blot (n = 6). (H) The FTL proteins measured by Western blot (n = 6). (I) The sections were labelled for DAPI (blue), NeuN (green), and FTL (red) (scale bar = 25 μm, n = 3). The results are presented as the mean ± SEM. Normal control group (Con), Chronic intermittent hypoxia group (CIH), Huperzine A-Liposomes group (HuA-LIP), Huperzine A group (HuA). *p < 0.05, **p < 0.01 vs Con group. #p < 0.05, ##p < 0.01 vs CIH group. $p < 0.05 vs HuA-LIP group.

FIGURE 7 Hepcidin-FPN1 involved in the iron deposition during CIH. (A) The hepcidin mRNA levels in heart tissue (n = 6). (B) The immunohistochemical staining of hepcidin protein (scale bar = 75 μm, n = 3). (C) The mean density of hepcidin content is shown in panel B (n = 3). (D) The immunofluorescence double label staining of FPN1 and hepcidin in heart tissue (scale bar = 25 μm, n = 3). The results are presented as the mean ± SEM. **p 

Figure 4 Hepcidin levels were elevated in CAG gastric tissue. (A) Hepcidin immunofluorescence staining in the gastric tissue of patients (Scale bar = 100 or 50 μm, n = 3). The bottom images were the larger images of the frame in the top images, and the arrows indicate the location of hepcidin-positive cells. (B) The mean density of hepcidin-positive cells in Panel (A). (C) Double immunofluorescence labeling of H+-K+-ATPase (green color) and hepcidin (red color) in the gastric tissue of CAG rats (Scale bar = 500 or 50 μm, n = 3). The right images (marked 1, 2) were the larger images of the frame in the Merge images. The data are presented as the mean ± SEM. ** p < 0.01 vs. NAG or Con groups. NAG: non-atrophic gastritis patients; CAG: chronic atrophic gastritis patients or rats; Con: the control group of rats.

Figure 4 FSGS rats showed iron metabolism disorder. (A) Prussian blue staining (scale bar: 100 μm). (B) Fe2+ results. (C) Western blot analysis of hepcidin, ferroportin and TFR. (D) Relative expression of hepcidin. (E) Relative expression of ferroportin. (F) Relative expression of TFR. *P 

Figure 5. Expression of iron transport-related gene and proteins. (A) Western blot analysis and quantitative analysis of (B) DMT1, (C) TFR, (D) Nu-SMAD1/5/8, (E) FPN, and (F) Hepcidin. (G) The mRNA relative expression of HAMP. (H) Iron deposits were assessed by Prussian blue staining; red arrows indicate iron deposits. Scale, 200 μm. LG = low-grain; HG = high-grain; DMT1 = divalent metal transporter; FPN = ferroportin; HAMP = hepcidin antimicrobial peptide; Nu-SMAD1/5/8 = nucleus-mothers against decapentaplegic homolog 1/5/8; TFRC = transferrin receptor. Values shown are mean ± SEM. Dots and squares represent the value of each sample. *P < 0.05, and **P < 0.01; ns = not significant.