rabbit polyclonal anti fsp1  (Proteintech)

Journal: Redox Biology

Article Title: A CoQ10 analog ameliorates cognitive impairment and early brain injury after subarachnoid hemorrhage by regulating ferroptosis and neuroinflammation

doi: 10.1016/j.redox.2025.103684

Figure Lengend Snippet: Erastin-induced ferroptosis promotes early brain injury after SAH A. The latency to eat food of aged mice. B. The time spent in the center of the aged mouse. C. The freezing time of aged mice. D. The levels of brain iron accumulation. E. The levels of brain lipid ROS. F. The mRNA levels of GPX4 and SLC7A11 were detected via RT‒qPCR. G. Ferroptosis-associated protein levels were detected via western blotting and quantitative analysis. H. Brain water content was determined via the standard wet-dry method. I. TUNEL staining of hippocampal tissues, and quantitative analysis (scale bar, 20 μm). J. Immunofluorescence staining of hippocampal tissues FSP1, and quantitative analysis (scale bar, 20 μm). The data are shown as the means ± SDs. ∗p < 0.05 and ∗∗p < 0.01.

Article Snippet: The primary antibodies utilized encompassed rabbit anti-NeuN polyclonal antibody (1:200, ab128886; Abcam, Cambridge, UK), rabbit anti-Iba1 monoclonal antibody (1:100, ab178847; Abcam), rabbit anti-GFAP antibody (1:100, abs119776; Absin), rabbit anti-FSP1 antibody (1:100, 20886-1-AP; Proteintech), and mouse anti-S100A10 antibody (1:100, 66227-1-Ig; Proteintech).

Techniques: Western Blot, TUNEL Assay, Staining, Immunofluorescence

Journal: Redox Biology

Article Title: A CoQ10 analog ameliorates cognitive impairment and early brain injury after subarachnoid hemorrhage by regulating ferroptosis and neuroinflammation

doi: 10.1016/j.redox.2025.103684

Figure Lengend Snippet: The neuroprotective effect of IDB is based on its anti-ferroptosis effects on neurons following SAH A-D. Hippocampal iron, ROS, MDA, and GSH levels in aged mice were detected via commercial kits and fluorescent probes. E-I. Western blotting images and quantitative data of relative protein level of FTH1, GPX4, SLC7A11, and ACSL4.J. Immunofluorescence staining of hippocampal tissues FSP1 (scale bar, 50 μm). K–N. The levels of Iron, MDA, ROS, and GSH in vitro experiments were detected via commercial kits and fluorescent probes.O. FSP1 immunofluorescence staining in vitro experiment (scale bar, 50 μm) P. Flow cytometry analysis of the intracellular ROS levels in vitro experiment.Q. Transmission electron microscopy images in vitro experiment (1 μm). The data are shown as the means ± SDs.

Article Snippet: The primary antibodies utilized encompassed rabbit anti-NeuN polyclonal antibody (1:200, ab128886; Abcam, Cambridge, UK), rabbit anti-Iba1 monoclonal antibody (1:100, ab178847; Abcam), rabbit anti-GFAP antibody (1:100, abs119776; Absin), rabbit anti-FSP1 antibody (1:100, 20886-1-AP; Proteintech), and mouse anti-S100A10 antibody (1:100, 66227-1-Ig; Proteintech).

Techniques: Western Blot, Immunofluorescence, Staining, In Vitro, Flow Cytometry, Transmission Assay, Electron Microscopy

Journal: Redox Biology

Article Title: A CoQ10 analog ameliorates cognitive impairment and early brain injury after subarachnoid hemorrhage by regulating ferroptosis and neuroinflammation

doi: 10.1016/j.redox.2025.103684

Figure Lengend Snippet: IDB treatment inhibited ferroptosis by interacting with FSP1 in aged SAH model mice A. Molecular docking. B. Protein-log and RMSD. C. Protein RMSF. D. Ligand‒protein contacts. E. Ligand torsion profile. F. Ligand properties. G-H. The levels of CoQ10H2 and CoQ10 were determined by ELISA.I. Representative Western blotting images of FSP1, β-action of Cellular Thermal Shift Assay (CESTA) in vitro experiment treated with or without IDB. The data are summarized as the mean ± SD.

Article Snippet: The primary antibodies utilized encompassed rabbit anti-NeuN polyclonal antibody (1:200, ab128886; Abcam, Cambridge, UK), rabbit anti-Iba1 monoclonal antibody (1:100, ab178847; Abcam), rabbit anti-GFAP antibody (1:100, abs119776; Absin), rabbit anti-FSP1 antibody (1:100, 20886-1-AP; Proteintech), and mouse anti-S100A10 antibody (1:100, 66227-1-Ig; Proteintech).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Thermal Shift Assay, In Vitro

Journal: Redox Biology

Article Title: A CoQ10 analog ameliorates cognitive impairment and early brain injury after subarachnoid hemorrhage by regulating ferroptosis and neuroinflammation

doi: 10.1016/j.redox.2025.103684

Figure Lengend Snippet: FSP1 deficiency reversed neuroprotection and the anti-ferroptosis effects of IDB A. LDH level in vitro experiment. B. Cell viability was measured by the WST-1 assay. C. Dead/Live staining (scale bar, 20 μm). D. Flow cytometry analysis of FITC-ROS in each group. E-G. The levels of MDA, GSH, and Iron in vitro experiment H. Images of ROS via DCFH-DA staining (scale bar, 20 μm).I. The levels of CoQ10H2/CoQ10 were determined by ELISA.J. Brain water content was determined via the standard wet-dry method.K. TUNEL staining of hippocampal tissues, and quantitative analysis (scale bar, 50 μm) The data are summarized as the mean ± SD.

Article Snippet: The primary antibodies utilized encompassed rabbit anti-NeuN polyclonal antibody (1:200, ab128886; Abcam, Cambridge, UK), rabbit anti-Iba1 monoclonal antibody (1:100, ab178847; Abcam), rabbit anti-GFAP antibody (1:100, abs119776; Absin), rabbit anti-FSP1 antibody (1:100, 20886-1-AP; Proteintech), and mouse anti-S100A10 antibody (1:100, 66227-1-Ig; Proteintech).

Techniques: In Vitro, WST-1 Assay, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, TUNEL Assay

Journal: Redox Biology

Article Title: A CoQ10 analog ameliorates cognitive impairment and early brain injury after subarachnoid hemorrhage by regulating ferroptosis and neuroinflammation

doi: 10.1016/j.redox.2025.103684

Figure Lengend Snippet: IDB regulates FSP1 protein stability through NMT-mediated N-myristoylation A. Western blot images and quantitative analysis of NMT1, NMT2, and β-actin. B. Western blot images and quantitative analysis of cytoplasmic FSP1, nuclear FSP1, membrane FSP1, histone H3, and β-actin. C. FSP1 immunofluorescence staining in vitro experiment. D-F. The levels of MDA, Iron, and GSH were detected via commercial kits in an in vitro experiment.G. Images of ROS via DCFH-DA staining.H. ROS levels detected by flow cytometry.I. Dead/Live staining (scale bar, 50 μm). The data are summarized as the mean ± SD.

Article Snippet: The primary antibodies utilized encompassed rabbit anti-NeuN polyclonal antibody (1:200, ab128886; Abcam, Cambridge, UK), rabbit anti-Iba1 monoclonal antibody (1:100, ab178847; Abcam), rabbit anti-GFAP antibody (1:100, abs119776; Absin), rabbit anti-FSP1 antibody (1:100, 20886-1-AP; Proteintech), and mouse anti-S100A10 antibody (1:100, 66227-1-Ig; Proteintech).

Techniques: Western Blot, Membrane, Immunofluorescence, Staining, In Vitro, Flow Cytometry

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 5 | IDA inhibits ferroptosis in retinal neurons via the AhR-ALDH1A3-FSP1 pathway. (a) Representative western blot images of f AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. IDA increased nAhR and ALDH1A3 levels, effects partially reversed by the AhR inhibitor (iAhR) but unaffected by ALDH1A3 or FSP1 inhibitors. FSP1 levels remained un- changed. (e–g) Quantification of oxidative stress markers. IDA reduced 4-HNE and MDA levels and increased GSH levels, effects negated by iAhR, iALDH1A3, or iFSP1. Data are mean ± SEM (n = 3). One-way ANOVA with Tukey's post hoc test for (d); unpaired t-test for (b, c, e–g). p values were adjusted for multiple testing by the Benjamini Hochberg method. **p < 0.01, ***p < 0.001 vs. control; ##p < 0.01, ###p < 0.001 vs. IDA group; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Western Blot, Control

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 6 | Neuroprotective effects of 7,8-DHF and the gut microbiota-IDA-AhR-ALDH1A3-FSP1 pathway on RGC survival and retinal function following ONC injury. (a) Retinal flat-mount images of RBPMS-labeled RGCs. Scale bar: 100 μm. (b) Representative PhNR traces showing retinal function. (c) Quantification of RGC survival (%). RGC density was reduced in the ONC group compared to the Sham group, partially preserved by 7,8-DHF, attenuated by antibiotics, and restored by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced RGC survival. (d) Quantification of PhNR amplitudes (μV). Retinal function decreased in the ONC group, was partially restored by 7,8-DHF, diminished by antibiotics, and recov- ered by IDA. AhR, ALDH1A3, and FSP1 inhibitors partially reduced the recovery. Data are mean ± SD (n = 6). Unpaired t-test was used except for Sham vs. ONC in (c), where Welch's t-test was applied; +++p < 0.001 vs. Sham; *p < 0.05, ***p < 0.001 vs. ONC; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. ONC + DHF; &&p < 0.01, &&&p < 0.001 vs. ONC + DHF + Abx.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Labeling

Journal: CNS neuroscience & therapeutics

Article Title: Oral 7,8-Dihydroxyflavone Protects Retinal Ganglion Cells by Modulating the Gut-Retina Axis and Inhibiting Ferroptosis via the Indoleacrylic Acid-AhR-ALDH1A3-FSP1 Pathway.

doi: 10.1111/cns.70442

Figure Lengend Snippet: FIGURE 7 | Effects of 7,8-DHF, gut microbiota disruption, and IDA on AhR-ALDH1A3-FSP1 pathway proteins and oxidative stress in retinal tissues following ONC injury. (a) Representative western blot images for AhR-ALDH1A3-FSP1 pathway proteins. (b–d) Quantification of nAhR, ALDH1A3, and FSP1 levels relative to β-actin. 7,8-DHF increased nAhR and ALDH1A3 levels, which were reduced by antibiotics and restored by IDA. AhR inhibitors decreased nAhR and ALDH1A3, while ALDH1A3 or FSP1 inhibition had no effect on nAhR. FSP1 levels remained unchanged across groups. (e–g) Oxidative stress markers: 7,8-DHF reduced 4-HNE and MDA while increasing GSH levels, effects reversed by antibiotics and restored by IDA. Inhibitors of AhR, ALDH1A3, and FSP1 partially reversed these effects, implicating the AhR-ALDH1A3-FSP1 pathway. Data are mean ± SEM (n = 3). p values were adjusted for multiple testing by the Benjamini Hochberg method. *p < 0.05, **p < 0.01 vs. ONC; #p < 0.05 vs. ONC + DHF; &p < 0.05, &&p < 0.01 vs. ONC + DHF + Abx; ns (above bars), p > 0.05 vs. ONC + DHF; ns (above horizontal lines), p > 0.05 among groups below the line.

Article Snippet: IDA's impact on ferroptosis proteins and AhR, ALDH1A3, and FSP1 inhibitors was examined in vitro and in vivo using rabbit anti- AhR (1:1000, #SAB4500725, Sigma), rabbit anti- ALDH1A3 (1:1000, #ABN427, Sigma), rabbit anti- FSP1 (1:1000, #A06541- 2, Boster), rabbit anti- GPX4 (1:1000, #SAB5700944, Sigma), rabbit anti- ACSL4 (1:2000, #SAB2100035, Sigma), rabbit anti- SLC7A11 (1:1000, #SAB5700735, Sigma), rabbit anti- GCH1 (1:1000, #PA5- 103865, Invitrogen), rabbit anti- FTH1 (1:500, #ZRB2695, Sigma), rabbit anti- DHODH (1:1000, #SAB2100574, Sigma), rabbit anti- 4- HNE (1:1000, #MA5- 27570, Invitrogen), and rabbit anti- β- actin (1:2000, #AF5003, Beyotime).

Techniques: Disruption, Western Blot, Inhibition