ACSL4/FACL4 Antibody - #DF12141

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 Fer-1 protects H9c2 cells against Herceptin-induced cell injury and ferroptosis. Fer-1 and DFO reversed the (A) Herceptin-induced reduction in cell viability, (B) Herceptin-induced decrease in GPX4 and SLC7A11 protein expression and Herceptin-induced increase in ACSL4 protein expression. Fer-1 and DFO reversed the Herceptin-induced (C) reduction in GSH content. (D) Fer-1 and DFO did not affect GSSG content. (E) Fer-1 and DFO reversed the Herceptin-induced reduction in the ratio of GSH/GSSG in H9c2 cells. Fer-1 and DFO reversed the Herceptin-induced increase in (F) intracellular and (G) mitochondrial iron levels in H9c2 cells. However, compared with DFO, the effects of Fer-1 were less potent. **P<0.01 and ***P<0.001 vs. NC. #P<0.05 and ##P<0.01 vs. Herceptin (10 µM). Fer-1, ferrostatin-1; GPX4, glutathione peroxidase 4; SLC7A11, recombinant solute carrier family 7 member 11; ACSL4, acyl-CoA synthetase long chain family member 4; GSH, reduced glutathione; GSSG, oxidized glutathione; DFO, deferoxamine; OD, optical density.

Figure 3. | Fer‑1 protects H9c2 cells against Herceptin‑induced cell injury and ferroptosis. Fer‑1 and DFO reversed the (A) Herceptin‑induced reduction in cell viability, (B) Herceptin‑induced decrease in GPX4 and SLC7A11 protein expression and Herceptin‑induced increase in ACSL4 protein expression.

FIGURE 5 The expression level of GPX4, ACSL4, and SLC7A11: (A) representative results of immunohistochemistry (IHC) and relative expression of GPX4; (B) representative results of IHC and relative expression of ACSL4; and (C) representative results of IHC and relative expression of SLC7A11. *p < 0.05 compared with the control group

Figure 4 DFO and EDA attenuated ferroptosis in EAP model. (a) The iron concentration of prostate lysates was determined by the commercial kit. Results were normalized to protein concentration. (b) The mRNA levels of ferroptosis biomarkers GPX4, SLC7A11, ACSL4, PTGS2, and DHODH relative to internal control were determined by the RT-PCR method. (c) The protein levels of ferroptosis biomarkers GPX4, SLC7A11, ACSL4, LPCAT3, and DHODH were determined by the western blot method. The relative quantification result of each band was performed relative to β-actin. Data was presented as mean ± SEM. #P < 0.05 versus the control group; ##P < 0.01 versus the control group; ###P < 0.001 versus the control group; ∗P < 0.05 versus the EAP group; ∗∗P < 0.01 versus the EAP group; ∗∗∗P < 0.001 versus the EAP group.