Ferritin Heavy Chain Antibody - #DF4828

FIGURE 1 Imatinib resistance is associated with ferroptosis activity in gastrointestinal stromal tumour (GIST). (A) Heatmap of differentially expressed genes (DEGs) in imatinib-naïve and -resistant GISTs from the GSE132542 dataset. (B) Gene ontology analysis of DEGs between imatinib-naïve and -resistant GISTs from the GSE132542 dataset (all absolute log2 fold change > .5, false discovery rate (FDR) < 10%). (C) Gene set enrichment analysis (GSEA) plots of negative regulation of cell death signalling pathway. (D) Western blotting analysis of caspase-3, cleaved caspase-3, caspase7, cleaved caspase-7, phosphorylated RIP1, total RIP1, phosphorylated RIP3, total RIP3, GSDME, cleaved GSDME, GSDMD, cleaved GSDMD, ferritin heavy chain (FTH1) and glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), FSP1 and dihydroorotate dehydrogenase (DHODH) expression in parental and imatinib-resistant (IR) GIST-882 cells treated with 882 20 µM imatinib for 24 h (for GIST-882 and 20 nM for GIST-T1, 24 h). Vinculin was included as a loading control. (E) Western blotting analysis of caspase-3, cleaved caspase-3, caspase7, cleaved caspase-7, phosphorylated RIP1, total RIP1, phosphorylated RIP3, total RIP3, GSDME, cleaved GSDME, GSDMD, cleaved GSDMD, FTH1 and GPX4, SLC7A11, FSP1 and DHODH expression in parental and IR GIST-T1 cells treated with 20 nM imatinib for 24 h. Vinculin was included as a loading control. (F) The expression of FTH1 and GPX4 in imatinib-sensitive and imatinib-resistant GIST specimens using immunohistochemistry (IHC) staining assay (Mann–Whitney U test). (G) Statistics of FTH1 expression in imatinib-sensitive (n = 30) and imatinib-resistant GIST specimens (n = 10) and percentage of GIST specimens with high or low FTH1 expression. (H) Statistics of GPX4 expression in imatinib-sensitive (n = 30) and imatinib-resistant GIST specimens (n = 10) and percentage of GIST specimens with high or low GPX4 expression. (I) Western blotting analysis showing the protein levels of FTH1, GPX4, SLC7A11, FSP1 and DHODH in six GIST specimens, including three samples from patients with imatinib-treated PD(Progressive Disease) and three samples from patients with imatinib-treated PR(Partial Response). Data represent the mean ± standard deviation (SD); *p < .05; **p < .01; ***p < .001. An unpaired t-test was used unless otherwise stated.

FIGURE 1 Imatinib resistance is associated with ferroptosis activity in gastrointestinal stromal tumour (GIST). (A) Heatmap of differentially expressed genes (DEGs) in imatinib-naïve and -resistant GISTs from the GSE132542 dataset. (B) Gene ontology analysis of DEGs between imatinib-naïve and -resistant GISTs from the GSE132542 dataset (all absolute log2 fold change > .5, false discovery rate (FDR) < 10%). (C) Gene set enrichment analysis (GSEA) plots of negative regulation of cell death signalling pathway. (D) Western blotting analysis of caspase-3, cleaved caspase-3, caspase7, cleaved caspase-7, phosphorylated RIP1, total RIP1, phosphorylated RIP3, total RIP3, GSDME, cleaved GSDME, GSDMD, cleaved GSDMD, ferritin heavy chain (FTH1) and glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), FSP1 and dihydroorotate dehydrogenase (DHODH) expression in parental and imatinib-resistant (IR) GIST-882 cells treated with 882 20 µM imatinib for 24 h (for GIST-882 and 20 nM for GIST-T1, 24 h). Vinculin was included as a loading control. (E) Western blotting analysis of caspase-3, cleaved caspase-3, caspase7, cleaved caspase-7, phosphorylated RIP1, total RIP1, phosphorylated RIP3, total RIP3, GSDME, cleaved GSDME, GSDMD, cleaved GSDMD, FTH1 and GPX4, SLC7A11, FSP1 and DHODH expression in parental and IR GIST-T1 cells treated with 20 nM imatinib for 24 h. Vinculin was included as a loading control. (F) The expression of FTH1 and GPX4 in imatinib-sensitive and imatinib-resistant GIST specimens using immunohistochemistry (IHC) staining assay (Mann–Whitney U test). (G) Statistics of FTH1 expression in imatinib-sensitive (n = 30) and imatinib-resistant GIST specimens (n = 10) and percentage of GIST specimens with high or low FTH1 expression. (H) Statistics of GPX4 expression in imatinib-sensitive (n = 30) and imatinib-resistant GIST specimens (n = 10) and percentage of GIST specimens with high or low GPX4 expression. (I) Western blotting analysis showing the protein levels of FTH1, GPX4, SLC7A11, FSP1 and DHODH in six GIST specimens, including three samples from patients with imatinib-treated PD(Progressive Disease) and three samples from patients with imatinib-treated PR(Partial Response). Data represent the mean ± standard deviation (SD); *p < .05; **p < .01; ***p < .001. An unpaired t-test was used unless otherwise stated.

FIGURE 4 Panax notoginseng saponins (PNS) suppresses ferroptosis by regulating iron metabolism and reducing lipid peroxidation. (a) Analysis of high-fat diet (HFD)-induced iron deposition in the aortic root of mice. The aortic root sections were stained with Prussian Blue and the iron content was quantified, n = 10. (b) Immunofluorescence analysis of GPX4 in aortic root sections from apoE−/− mice, and mean fluorescence intensity was quantified, n = 10. (c–e) Serum MDA, SOD and CAT levels of apoE−/− mice were measured by Elisa assay kits, n = 10. (f) Quantification of expression of genes encoding for lipid peroxide clearance (Gpx4 and Slc7A11), peroxide generation (Acsl4 and Lpcat3) and iron metabolism (Fth), with qRT-PCR, in peritoneal macrophages from apoE−/− mice, n = 6. (g) Quantification of NCOA4, SLC7A11, FtH and GPX4 protein expression via western blotting in peritoneal macrophages from apoE−/− mice, n = 6. Data shown are means ± SEM (n = 6). *P < 0.05, significantly different from control (Ctrl).