fth1 antibody  (Cell Signaling Technology Inc)

Journal: Redox Biology

Article Title: ACOD-itaconate in macrophage attenuates oxidative stress and inflammation in benign airway stenosis by upregulating and transferring FTH1

doi: 10.1016/j.redox.2026.104133

Figure Lengend Snippet: ACOD1/itaconate axis upregulates FTH1 to mitigate oxidative stress in macrophages. A. Re-clustering of the Mc4 macrophage subpopulation based on AcoD1 expression identifies ACOD1 + and ACOD1 - subsets. B. KEGG pathway enrichment analysis shows the necroptosis pathway is most significantly enriched in ACOD1 + macrophages compared to ACOD1 - subsets. Top 5 enriched pathways shown. C. Differential gene expression analysis within the necroptosis pathway identifies FTH1 as the most upregulated gene in ACOD1 + macrophages. D. Quantitative comparison of FTH1 expression levels in scRNA-seq data between normal trachea group and granulation group. E, F. Representative images of FTH1 immunohistochemical staining and quantitative of immunoreactive area in normal trachea group and granulation group. Scale bars indicate 200 μm and 50 μm (n = 3 sample per group). G, H. Western blot analysis showing the levels of ACOD1 in the trachea of control group, BAS group and BAS+4-OI group (n = 3 mice per group) at 24 h. I, J. Representative tracheal immunofluorescence images and mean fluorescence intensity of FTH1(green) in control group, BAS group and BAS+4-OI group (n = 5 mice per group) at 24 h. Scale bars indicate 200 μm and 50 μm. K. Representative immunofluorescence images of CD68(red), FTH1(green) and ACOD1(yellow)in normal trachea group and granulation group. Scale bars indicate 200 μm and 50 μm. L. Representative tracheal immunofluorescence images of F4/80(red), FTH1(green) and ACOD1(yellow)in control group and BAS group. Scale bars indicate 200 μm and 50 μm. M, N. Western blot analysis in macrophages reveals that 4-OI rescues LPS-induced FTH1 downregulation (n = 3 independent experiments). O, P. Representative immunofluorescence images and mean fluorescence intensity of ROS (red) in 4-OI reduces LPS-induced 4-HNE in controls, but not in FTH1-knockdown macrophages. (n = 3 independent experiments). Scale bars indicate 50 μm. Q, R. Western blot and quantitative analysis of FTH1 in macrophages after transfection with siNRF2 and treated or not with 4-OI, LPS (n = 3 independent experiments). Data are presented as the mean ± SEM. Two-sided student's T-test were used in D,F. One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in H, J. Two-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in N, P, R.

Article Snippet: The IP group received FTH1 antibody (#4393, Cell Signaling Technology, 1:50), while the IgG control group received rabbit IgG antibody.

Techniques: Expressing, Gene Expression, Comparison, Immunohistochemical staining, Staining, Western Blot, Control, Immunofluorescence, Fluorescence, Knockdown, Transfection

Journal: Redox Biology

Article Title: ACOD-itaconate in macrophage attenuates oxidative stress and inflammation in benign airway stenosis by upregulating and transferring FTH1

doi: 10.1016/j.redox.2026.104133

Figure Lengend Snippet: 4-OI promotes exosomal FTH1 secretion from macrophages and induces fibroblast ferroptosis. A. ELISA detects elevated FTH1 secretion in supernatants of macrophages treated with LPS + 4-OI, but not 4-OI alone (n = 3 independent experiments). B. Representative TEM images of exosomes isolated from LPS+4-OI treated macrophages. Scale bars indicate 500 nm and 200 nm. C. Representative NTA of exosomes isolated from LPS+4-OI treated macrophages. D. Representative immunoblot shows exosome markers (TSG101, CD63, and CD81) and endoplasmic reticulum marker (Calnexin). E. Schematic diagram of exosome uptake experiment. F. Representative immunofluorescence images of DiD (red) and F-actin (green) in two fibroblasts group. Scale bars indicate 20 μm. G. Representative immunoblot shows FTH1 and CD63 in two EXO group. H,I. Representative immunofluorescence images and mean fluorescence intensity of FTH1 (red) in three fibroblasts group (n = 3 independent experiments). Scale bars indicate 50 μm. J. Experimental scheme: Macrophage-fibroblast co-culture system. K,L. Representative immunofluorescence images and mean fluorescence show co-culture with macrophages treated with LPS+4-OI significantly enhanced FTH1 internalization in fibroblasts. This effect was abolished when macrophages were subjected to FTH1 knockdown (LPS+4-OI + siFTH1), and was restored upon addition of recombinant FTH1 (LPS+4-OI + siFTH1+rFTH1) (n = 3 independent experiments). Scale bars indicate 50 μm. M,N. Representative immunofluorescence images and mean fluorescence of PGSK in fibroblasts after co-culture with the indicated macrophage groups: control, LPS+4-OI, LPS+4-OI + siFTH1, and LPS+4-OI + siFTH1+rFTH1 (n = 3 independent experiments). Scale bars indicate 100 μm. O,P. Representative immunofluorescence images and Mean fluorescence intensity of BODIPY C11(red) and oxidized form(green) in two group(n = 3 independent experiments) Scale bars indicate 50 μm. Q. Representative TEM images of differently treated fibroblasts group. Scale bars indicate 2 μm and 500 nm. R,S. Representative immunofluorescence images and mean fluorescence intensity of 4HNE (red) in three fibroblasts group (n = 3 independent experiments). Scale bars indicate 50 μm. Data are presented as the mean ± SEM. Two-sided student's T-test were used in P. One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in A,I,L,N,S .

Article Snippet: The IP group received FTH1 antibody (#4393, Cell Signaling Technology, 1:50), while the IgG control group received rabbit IgG antibody.

Techniques: Enzyme-linked Immunosorbent Assay, Isolation, Western Blot, Marker, Immunofluorescence, Fluorescence, Co-Culture Assay, Knockdown, Recombinant, Control, Comparison

Journal: Redox Biology

Article Title: ACOD-itaconate in macrophage attenuates oxidative stress and inflammation in benign airway stenosis by upregulating and transferring FTH1

doi: 10.1016/j.redox.2026.104133

Figure Lengend Snippet: Macrophage-derived FTH1 binds fibroblast SCARA5 to inhibit fibrosis. A. Re-clustering of fibroblasts from single-cell RNA-seq data identifies distinct subpopulations in human BAS granulation tissue. B. Pseudotime trajectory analysis reveals fibroblast differentiation converging on pro-fibrotic Cluster 2. C. CellChat interaction analysis predicts FTH1 (from ACOD1 + macrophages) binding to SCARA5 (on Cluster 2 fibroblasts) as the top ligand-receptor pair. D. Representative immunofluorescence images of FTH1(green), SCARA5(yellow) and Vimentin (red) in normal trachea group and granulation group. Scale bars indicate 50 μm and 20 μm. E. Co-immunoprecipitation (Co-IP) confirms FTH1 binding to SCARA5 in fibroblasts co-cultured with LPS+4-OI-treated macrophages. F,G. Representative tracheal immunofluorescence images of FTH1 (green), GPX4(yellow), SCARA5(red) and VIMENTIN(pink) in mouse trachea and quantitative of mean fluorescence of FTH1 and GPX4 in the control, BAS and BAS+4-OI group(n = 3 mice per group). Scale bars indicate 200 μm and 50 μm. H. Schematic diagram of the experimental design for 4-OI and Anti -SCARA5 treatment. I,J. Representative immunofluorescence images and mean fluorescence intensity of αSMA (red) and COL1(green) in four group: Control, BAS,BAS+4-OI,BAS+4-OI + Anti -SCARA5 (n = 3 mice per group). Scale bars indicate 200 μm and 50 μm. Data are presented as the mean ± SEM. One-way ANOVA analysis followed by Tukey post hoc multi-comparison test was used in G, J.

Article Snippet: The IP group received FTH1 antibody (#4393, Cell Signaling Technology, 1:50), while the IgG control group received rabbit IgG antibody.

Techniques: Derivative Assay, Single Cell, RNA Sequencing, Binding Assay, Immunofluorescence, Immunoprecipitation, Co-Immunoprecipitation Assay, Cell Culture, Fluorescence, Control, Comparison

Journal: Redox Biology

Article Title: ACOD-itaconate in macrophage attenuates oxidative stress and inflammation in benign airway stenosis by upregulating and transferring FTH1

doi: 10.1016/j.redox.2026.104133

Figure Lengend Snippet: Schematic diagram of mechanism of the ACOD1/itaconate axis in regulating airway inflammation and fibrosis. During the inflammatory stage, macrophages upregulate ACOD1 expression, catalyzing itaconate production. Itaconate and its derivative 4-OI activate the NRF2 signaling pathway by promoting NRF2 nuclear translocation. Nuclear NRF2 induces FTH1 transcription, leading to intracellular FTH1 protein accumulation to suppressing inflammation and attenuating oxidative stress. In fibrotic repair, FTH1 packaged into exosomes and secreted. These exosomes are taken up by fibroblasts via bind to SCARA5, leading to FTH1 delivery and subsequent induction of ferroptosis.

Article Snippet: The IP group received FTH1 antibody (#4393, Cell Signaling Technology, 1:50), while the IgG control group received rabbit IgG antibody.

Techniques: Expressing, Translocation Assay