intracellular ferrous iron fe2 levels  (Elabscience Biotechnology)

Journal: Advanced Science

Article Title: Xanthatin Targets CISD1 to Drive Ferroptosis and Mitophagy as a Dual Anticancer Strategy in Triple‐Negative Breast Cancer

doi: 10.1002/advs.202520051

Figure Lengend Snippet: Xanthatin induces ferroptosis in TNBC cells. (A) KEGG enrichment analysis of DEGs in MDA‐MB‐231 cells treated with xanthatin for 48 h revealed significant enrichment in ferroptosis‐related pathways. (B) Western blot analysis showing the expression levels of ferroptosis‐associated proteins SLC7A11, FTH1, TfR1, and GPX4 in MDA‐MB‐231 and MDA‐MB‐468 cells after treatment with xanthatin for 48 h. (C) Flow cytometry analysis of ROS levels in MDA‐MB‐231 and MDA‐MB‐468 cells treated with xanthatin for 48 h. (D) MDA content in TNBC cells after xanthatin treatment for 48 h. (E) GSH levels in TNBC cells treated with xanthatin for 48 h. (F) Intracellular Fe 2 + levels determined by flow cytometry in TNBC cells after xanthatin treatment for 48 h. (G) Representative images of JC‐1 staining showing loss of mitochondrial membrane potential in MDA‐MB‐231 and MDA‐MB‐468 cells after xanthatin treatment for 48 h. Scale bar = 100 µm. Bars, SDs; * 0.01< p < 0.05, ** 0.001< p < 0.01, and *** p < 0.001.

Article Snippet: Intracellular ferrous ion (Fe 2 + ) levels were quantitatively assessed using the Fe 2 + ‐specific fluorescent probe FerroOrange (HY‐K0322, MedChemExpress).

Techniques: Western Blot, Expressing, Flow Cytometry, Staining, Membrane

Journal: Advanced Science

Article Title: Xanthatin Targets CISD1 to Drive Ferroptosis and Mitophagy as a Dual Anticancer Strategy in Triple‐Negative Breast Cancer

doi: 10.1002/advs.202520051

Figure Lengend Snippet: Xanthatin‐induced mitophagy is activated in response to ferroptotic stress. (A) Immunofluorescence staining showing colocalization of LC3 and mitochondria in MDA‐MB‐231 and MDA‐MB‐468 cells treated with xanthatin for 48 h, with or without DFO pretreatment (20 µ m , 12 h). Scale bar = 20 µm. (B) Western blot analysis of mitophagy‐related proteins Parkin, TOM20, and LC3 in MDA‐MB‐231 and MDA‐MB‐468 cells treated with xanthatin in the presence or absence of 20 µM DFO. (C‐E) Flow cytometric analysis of intracellular ROS, Fe 2 + and 4‐HNE levels in MDA‐MB‐231 and MDA‐MB‐468 cells treated with xanthatin for 48 h, with or without 1 mM 3‐MA pretreatment.

Article Snippet: Intracellular ferrous ion (Fe 2 + ) levels were quantitatively assessed using the Fe 2 + ‐specific fluorescent probe FerroOrange (HY‐K0322, MedChemExpress).

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Advanced Science

Article Title: Xanthatin Targets CISD1 to Drive Ferroptosis and Mitophagy as a Dual Anticancer Strategy in Triple‐Negative Breast Cancer

doi: 10.1002/advs.202520051

Figure Lengend Snippet: The anticancer activity of xanthatin relies on CISD1. (A‐B) CCK‐8 assay (A) and EdU incorporation assay (B) in MDA‐MB‐231 and MDA‐MB‐468 stable knockdown cell lines (shCISD1#1, shCISD1#2) and control cells (shNC) treated with DMSO or xanthatin for 48 h. Scale bar for EdU = 100 µm. (C‐D) Flow cytometry analysis of ROS levels (C) and intracellular Fe 2 + levels (D) in shNC, shCISD1#1, and shCISD1#2 stable knockdown cell lines treated with DMSO or xanthatin for 48 h. (E‐F) Biochemical detection of MDA levels (E) and GSH levels (F) in shNC, shCISD1#1, and shCISD1#2 stable knockdown cell lines treated with DMSO or xanthatin for 48 h. Bars, SDs; * 0.01< p < 0.05, ** 0.001< p < 0.01, and *** p < 0.001. n.s., no significance.

Article Snippet: Intracellular ferrous ion (Fe 2 + ) levels were quantitatively assessed using the Fe 2 + ‐specific fluorescent probe FerroOrange (HY‐K0322, MedChemExpress).

Techniques: Activity Assay, CCK-8 Assay, Knockdown, Control, Flow Cytometry

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Genetic Regulation of Alternative Polyadenylation Provides Novel Insights into Molecular Mechanisms Underlying Non-small Cell Lung Cancer.

doi: 10.1002/advs.202502008

Figure Lengend Snippet: Figure 6. Risk T allele at LYRM4 suppressing ferroptosis activity to promote malignant phenotypes of NSCLC cells. A) Schematic of LYRM4 interac- tor identification through proteomic screening by mass spectrometry in A549 cells. This figure was created with BioRender (www.biorender.com). B) Immunoprecipitation followed by proteomic screening showed increased binding of NFS1 in LYRM4[T] overexpression group compared to LYRM4[G] overexpression group. Horizontal line indicates the P < 0.05 cutoff. Vertical lines mark the fold change > 1.1 or < 0.9. C) Coimmunoprecipitation followed by western blotting analysis confirmed the interaction between NFS1 and LYRM4 isoforms. D–G) Cell viability D), Fe2+ levels E), MDA levels F), and

Article Snippet: Cellular Iron Detection: Intracellular ferrous iron (Fe2+) levels were detected with the Cell Ferrous Iron Colorimetric Assay Kit (E-BC-K881-M, Elabscience, China) and the absorbance at 593 nm was calculated as the intracellular Fe2+ levels.

Techniques: Activity Assay, Mass Spectrometry, Immunoprecipitation, Binding Assay, Over Expression, Western Blot