Journal: Viruses

Article Title: Infectious Spleen and Kidney Necrosis Virus Triggers Ferroptosis in CPB Cells to Enhance Virus Replication.

doi: 10.3390/v17050713

Figure Lengend Snippet: Figure 3. ISKNV infection induces ferroptosis in CPB cells. (A) Transmission electron microscopy of CPB cells treated with DMSO (72 h), erastin (10 µmol/L, 72 h), and ISKNV (100 MOI, 24 h, 48 h, 72 h). Scale bars = 1 µm. (B) Analysis of Fe2+ levels in CPB cells after treatment with ISKNV (100 MOI) using the fluorescent probe FerroOrange by laser scanning confocal microscopy. Scale bars = 20 µm. (C) Quantitative analysis of the mean fluorescence intensity of (B) using Image J. (D) Analysis of intracellular ROS levels using DCFH-DA staining, and laser scanning confocal microscopy of CPB cells treated with ISKNV (100 MOI) for 24–72 h. Scale bars = 10 µm. (E) Quantitative analysis of the mean fluorescence intensity of (D) using Image J. (F–H) Detection of Fe2+, ROS, and MDA levels in cell lysates treated with ISKNV (100 MOI) for 24–72 h by microplate reader. * p < 0.05, ** p < 0.01, and *** p < 0.001, with p > 0.05 considered not significant (ns).

Article Snippet: The Fe2+ content of the cells was detected by laser scanning confocal microscopy and a microplate reader using the fluorescent probe FerroOrange (Elabscience, E-BC-F101, Wuhan, China).

Techniques: Infection, Transmission Assay, Electron Microscopy, Confocal Microscopy, Fluorescence, Staining

Journal: Discover Oncology

Article Title: Novel N 6 -methyladenosine (m 6 A) writer METTL16 promotes the cervical cancer tumorigenesis by targeting FTH1-dependent ferroptosis

doi: 10.1007/s12672-026-04403-8

Figure Lengend Snippet: METTL16 inhibited the ferroptosis of cervical cancer cells. A Transmission electron microscopy (TEM) was performed to detect the cellular substructures transformation in cervical cancer cells with Erastin (10 µmol/L). HeLa cells were transfected with overexpression of METTL16 plasmids (ov-NC, ov-METTL16). CaSki cells were transfected with silencing of METTL16 plasmids (sh-NC, sh-METTL16-1, sh-METTL16-2). B The Fe 2+ level in cervical cancer cells was tested by Iron Colorimetric Assay Kit. C MDA was tested in cervical cancer cells. D GSH was tested in cervical cancer cells. E , F FerroOrange fluorescent assay was performed to test the Fe 2+ in cancer cells. * p < 0.05, ** p < 0.01

Article Snippet: FerroOrange fluorescent assay was performed to test the Fe 2+ in cancer cells FerroOrange fluorescent probe (Servicebio, Cat. G1727).

Techniques: Transmission Assay, Electron Microscopy, Transformation Assay, Transfection, Over Expression, Colorimetric Assay, Fluorescence

Journal: Science Advances

Article Title: Macrophage-targeted Mms6 mRNA-lipid nanoparticles promote locomotor functional recovery after traumatic spinal cord injury in mice

doi: 10.1126/sciadv.ads2295

Figure Lengend Snippet: ( A ) Schematics of the experimental workflow. SCI mice were received a single injection of Mms6 mRNA-LNPs. ( B and C ) Basso Mouse Scale (BMS) scores and subscores were assessed in SCI mice ( n = 6 mice per group). * P < 0.05 and ** P < 0.01. ( D ) Representative footprint images at 28 dpi of SCI. The forelimbs and hindlimbs were painted in black and red, respectively. ( E ) Evaluation of stride length using footprint images ( n = 6 mice per group). ** P < 0.01 versus Sham group; ## P < 0.01 versus empty LNP group; && P < 0.01 versus Mms6 mRNA-LNP (0.5 mg/kg) group; $ P < 0.05 versus Mms6 mRNA-LNP (1 mg/kg) group. ( F ) Statistical analysis of flow cytometry results showing the percentage of FerroOrange positive Mφ ( n = 3 mice per group). ** P < 0.01 versus empty LNP group. ( G ) Statistical analysis of flow cytometry results showing the percentage of Liperfluo positive Mφ ( n = 3 mice per group). ** P < 0.01 versus empty LNP group. dpi, days post-injury.

Article Snippet: Intracellular ferrous ions and lipid peroxides in macrophages were detected using FerroOrange and Liperfluo fluorescent probes, respectively (Dojindo, Shanghai, China).

Techniques: Injection, Flow Cytometry

Journal: Journal of Extracellular Vesicles

Article Title: Transfer of inflammatory mitochondria via extracellular vesicles from M1 macrophages induces ferroptosis of pancreatic beta cells in acute pancreatitis

doi: 10.1002/jev2.12410

Figure Lengend Snippet: Assessment of mitochondrial reactive oxygen species (mito ROS), ferrous ion (Fe2+) concentration, lipid peroxidation content, and mitochondrial morphology in Beta TC‐6 cells following various treatment protocols. (a) Representative images of mito ROS detection in Beta TC‐6 cells after treatment with M1 mitochondria, M1‐EVs, or a combination of both along with Fer‐1 ( n = 3). (b) Representative images of assessment of Fe2+ level in Beta TC‐6 cells after different treatment using FerroOrange fluorescence probe ( n = 3). (c) Representative images of detection of lipid peroxidation content in Beta TC‐6 cells after different treatments using mitoPeDPP fluorescence probe ( n = 3). (d) Representative images of mitochondrial morphology and quantitative evaluation of mitochondria in Beta TC‐6 cells following various treatments, utilizing a transmission electron microscope ( n = 10). Red triangle, abnormal mitochondria treated by mitochondria of M1 macrophages; Yellow triangle, abnormal mitochondria treated by EVs derived from M1 macrophages; Purple triangle, abnormal mitochondria treated by mitochondria of M1 macrophages combined with Fer‐1; Blue triangle, abnormal mitochondria treated by EVs derived from M1 macrophages combine with Fer‐1. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The intracellular Fe 2+ and mitochondrial lipid peroxidation of TC‐6 cells after different treatments were assessed using a FerroOrange fluorescent probe (DOJINDO, F374, Japan) and MitoPeDPP fluorescent probe (DOJINDO, M466, Japan), respectively, following the manufacturer's instructions.

Techniques: Concentration Assay, Fluorescence, Transmission Assay, Microscopy, Derivative Assay