SLC40A1 Antibody - #DF13561

Figure 3 EPO regulates the expression of ferroptotic biomarkers after SCI. (A) Western blot of the indicated proteins in injured tissues from the treatment groups. (B–H) Quantitative analysis of Tfr, Fpn, Fth, Acsl4 and 4-Hne protein expression at 14 dpi. Gapdh was used as the reference protein. (I, J) Quantitative reverse transcription polymerase chain reaction results of xCT and Gpx4 mRNA extracted from injured tissue at 7 dpi. β-Actin mRNA was used as the reference gene. (K) Quantitative analysis of reduced GSH from injured tissue at 7 dpi. All data are expressed as the mean ± SD (n = 5 in each group). *P < 0.05, **P < 0.01, ***P < 0.001 (one-way analysis of variance followed by Dunnett’s multiple comparisons test). 4-Hne: 4-Hydroxynonenal; Acsl4: acyl-coenzyme A synthetase long chain family member 4; dpi: day(s) post injury; EPO: erythropoietin; Fpn: ferroportin or solute carrier family 40 member 1; Fth: ferritin heavy chain; Gapdh: glyceraldehyde-3-phosphate dehydrogenase; Gpx4: glutathione peroxidase 4; GSH: glutathione; ns: not significant; SCI: spinal cord injury; Tfr: transferrin receptor; xCT: the solute carrier family 7 member 11.

Figure 3 Hepatic genes expression involved in iron metabolism. (a) Hepatic mRNA expression of iron metabolism (n = 6). (b) Protein content of ZIP14 in the liver (n = 6). (c) Protein content of DMT1 in the liver (n = 6). (d) Protein content of FTL in the liver (n = 6). (e) Protein content of FPN in the liver (n = 6). (f) Hepatic mRNA expression of HAMP and the upstream regulators (n = 6). Values are means ± SEM. # p < 0.05 and ## p < 0.01 compared with the CON group. * p < 0.05 and ** p < 0.01 compared with the HF group.

Fig. 2. Idebenone inhibited H2O2-induced ferroptosis. (A) The effect of deferoxamine (DFO, 100 μM) on the viability of H9c2 cells treated with 100 μM H2O2 for 4 h as detected by CCK8 assay (n = 3). (B) The GPX4, FTH1 and FPN1 protein expressions were detected via western blotting. (C-E) Quantitative analysis of GPX4, FTH1and FPN1 protein expressions (n = 3). (F) ROS level in H9c2 cells was assessed by DCFH-DA probe (scale bar:50 μm). (G) Quantitative analysis of ROS level in H9c2 cells (n = 3). (H) Quantitative analysis of MDA level in H9c2 cells (n = 3). (I) Quantitative analysis of ATP level in H9c2 cells (n = 3). (J) Representative images of JC-1 fluorescence (scale bar:50 μm). (K) Quantitative analysis of MMP level (n = 3). (L) Representative images of intracellular Fe2+ levels detected by FerroOrange (scale bar:100 μm). (M) Quantitative analysis of Fe2+ level (n = 3). The data are expressed as means ± SD. *P < 0.05, **P < 0.01 and && P < 0.01 compared with control group; #P < 0.05, ##P < 0.01 compared with H2O2 group.