Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Celluar study design. (B) After SIRT1 overexpression, western blotting was performed on H9c2 cells to determine PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 levels, and grayscale values were analyzed, with β-actin serving as a protein loading control. Intracellular levels of FFA (C) and A-CoA (D) after SIRT1 overexpression in H9c2 cells. (E) Intracellular ATP levels in H9c2 cells after SIRT1 overexpression. (F) Intracellular GSH/GSSG levels in H9c2 cells after SIRT1 overexpression. (G) Representative images of TMRE staining and quantitative analysis of fluorescence (I) in H9c2 cells after SIRT1 overexpression. Scale bar = 100 μm. (H) Representative images of DCFH-DA staining and quantitative analysis of fluorescence (J) in H9c2 cells after SIRT1 overexpression. Scale bar = 100 μm. (K) CCK-8 assays were employed to assess the viability of H9c2 cells after SIRT1 overexpression. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. Hypoxia.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Over Expression, Western Blot, Control, Staining, Fluorescence, CCK-8 Assay

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Celluar study design. (B) After overexpression of SIRT1, PPARα was inhibited by GW6471 in H9c2 cells. Western blotting was conducted to measure the levels of PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 in H9c2 cardiomyocytes, and grayscale values were analyzed. β-actin was used as a protein loading control. Intracellular levels of FFA (C), A-CoA (D), ATP (E), and GSH/GSSG (F) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. (G) Representative images of TMRE staining and quantitative analysis of fluorescence (I) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. Scale bar = 100 μm. (H) Representative images of DCFH-DA staining and quantitative analysis of fluorescence (J) in H9c2 cells after SIRT1 overexpression and PPARα inhibition. Scale bar = 100 μm. (K) CCK-8 assays were employed to assess the viability of H9c2 cells after SIRT1 overexpression and PPARα inhibition. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. Hypoxia, &P < 0.05, &&P < 0.01 vs. Hypoxia+SIRT1 OE .

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Over Expression, Western Blot, Control, Inhibition, Staining, Fluorescence, CCK-8 Assay

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: (A) Animal study design. (B) Echocardiography in rats. (C-E) RSV decreased LVPWd and LVPWs levels and increased LVEDV, LVESV, SV, and CO levels of the heart in the HH test. (F) RSV decreased the cardiac coefficient compared with the HH group. (G) Transmission electron microscopy was utilized to visualize the left ventricular myocardium ultrastructure in rats (8,000×). Scale bar = 1 μm. (H, I) A detailed analysis was conducted to determine the number of mitochondria per unit area and the average mitochondrial size. Myocardial FFA (J), A-CoA (K), and ATP (L) levels in rats after treatment with RSV. (M) DHE staining of rat myocardial tissue (40×) after RSV treatment and quantitative analysis of fluorescence (O). (N) Western blotting was performed on rat myocardial tissue extracts to determine SIRT1, PGC1α, PPARα, PDK4, CPT1, GPX4, and SLC7A11 levels, and grayscale values were analyzed (Q), with β-actin serving as a protein loading control. (P) Myocardial GSH/GSSG levels in rats after treatment with RSV. Statistical significance was indicated as *P < 0.05, **P < 0.01 vs. Control, #P < 0.05, ##P < 0.01 vs. HH.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques: Transmission Assay, Electron Microscopy, Staining, Fluorescence, Western Blot, Control

Journal: bioRxiv

Article Title: Regulation of fatty acid oxidation involved in high altitude hypoxia induced cardiomyocyte ferroptosis via the SIRT1-PPARα-GPX4 signaling pathway

doi: 10.1101/2025.01.20.633884

Figure Lengend Snippet: Under conditions of HH exposure, the downregulation of SIRT1 triggers FAO and ferroptosis through the PPARα-PDK4/CPT1-GPX4/SLC7A11 pathway. This results in mitochondrial dysfunction, ultimately contributing to the development of cardiac impairment, as depicted in the pathway map.

Article Snippet: Antibodies against PPARα, pyruvate dehydrogenase kinase 4 (PDK4), carnitine palmitoyltransferase 1 (CPT1), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC1α), SIRT1, GPX4, SLC7A11, and β-actin were obtained from Proteintech (Wuhan, China) and Abcam (Cambridge, UK).

Techniques:

Journal: Apoptosis

Article Title: Targeting FAR1 to inhibit intestinal macrophage ferroptosis: repurposing irinotecan as a novel sepsis therapy

doi: 10.1007/s10495-026-02419-7

Figure Lengend Snippet: Genetic inhibition of FAR1 mitigates intestinal macrophage ferroptosis and improves sepsis outcomes. A Stacked bar plot illustrating the relative proportions of 22 infiltrating immune cell types in sepsis patient samples estimated with the CIBERSORT algorithm. Each column represents an individual sample. Representative fluorescence images of B intracellular Fe 2+ (FerroOrange; scale bar = 25 μm) and lipid peroxide (Liperfluo; scale bar = 25 μm) 24 h after the Control or LPS (500 ng/mL) treatment with or without shFAR1. C , D Western blot and quantitative analysis of ACSL4 and GPX4 expression ( n = 3 per group). E Kaplan‒Meier survival analysis of mice after 72 h of sham surgery or sepsis induction with or without pretreatment with siFAR1 or siNC (10 nmol/20 g, i.p.) ( n = 16 per group). F Representative images of H&E staining of intestinal tissue (scale bars: 200 μm). G Representative TEM image of mitochondria in intestinal macrophages (scale bar = 1 μm). Flow cytometry analysis of intestinal macrophages was conducted, and the levels of H intracellular Fe 2+ (FerroOrange) and I lipid peroxide (Liperfluo) were quantified in mice 24 h post-surgery, with or without siFAR1 or siNC pretreatment (10 nmol/20 g, i.p.) ( n = 3 per group). The data are presented as the mean ± standard deviation; ** p < 0.01 versus the Control + shNC/Sham + siNC group; ## p < 0.01 versus the LPS + shNC/Sepsis + siNC group. Abbreviations: shNC, short hairpin RNA negative control; shFAR1, short hairpin RNA targeting FAR1; siNC, small interfering RNA negative control; siFAR1, small interfering RNA targeting FAR1; LPS, lipopolysaccharide; H&E, hematoxylin and eosin; TEM, transmission electron microscopy

Article Snippet: Primary antibodies against the following proteins were used: NRF2 (1:1000; A21176), ACSL4 (1:1000; A2041) and GPX4 (1:1000; A11243) from ABclonal (Wuhan, China); FAR1 (1:1000; NBP1-89847) from Novus Biologicals (Centennial, CO, USA); GSDMD-NT(1:1000; 10137T), p-MLKL(1:1000; 37333T), MLKL(1:1000; 37705T), p-p65(1:1000; 3033T), and p65(1:1000; 8242T) from Cell Signaling Technology (Danvers, MA, USA); cleaved Caspase-3(1:1000; 68773-1-Ig) and β-actin (1:5000; 20536-1-AP) from Proteintech (Wuhan, China).

Techniques: Inhibition, Fluorescence, Control, Western Blot, Expressing, Staining, Flow Cytometry, Standard Deviation, shRNA, Negative Control, Small Interfering RNA, Transmission Assay, Electron Microscopy

Journal: Apoptosis

Article Title: Targeting FAR1 to inhibit intestinal macrophage ferroptosis: repurposing irinotecan as a novel sepsis therapy

doi: 10.1007/s10495-026-02419-7

Figure Lengend Snippet: Irinotecan attenuates macrophage ferroptosis in vitro. A Representative TEM images of mitochondria in RAW264.7 cells (scale bar = 1 μm). Flow cytometry analysis of B intracellular Fe 2+ (FerroOrange), D lipid peroxide (Liperfluo), and E ROS (DCFH-DA) levels in RAW264.7 cells 24 h after the Control or LPS (500 ng/mL) treatment with or without irinotecan pretreatment (1 µM). C Representative fluorescence images of intracellular Fe 2+ (FerroOrange; scale bar = 25 μm), lipid peroxides (Liperfluo; scale bar = 25 μm), and ROS (DCFH-DA; scale bar = 25 μm). ELISA quantification of F) MDA levels, G the GSH/GSSG ratio, H TNF-α levels, and I) IL-6 levels in cell supernatants ( n = 8 per group). J Western blot of NRF2, ACSL4, FAR1, and GPX4 expression. K Quantitative analysis of the protein levels in panel J ( n = 3 per group). The data are presented as the mean ± standard deviation; ** p < 0.01 versus the Control group; ## p < 0.01 versus the LPS group. Ir, irinotecan; LPS, lipopolysaccharide; TEM, transmission electron microscopy; MDA, malondialdehyde; ROS, reactive oxygen species; GSH/GSSG, glutathione/oxidized glutathione ratio

Article Snippet: Primary antibodies against the following proteins were used: NRF2 (1:1000; A21176), ACSL4 (1:1000; A2041) and GPX4 (1:1000; A11243) from ABclonal (Wuhan, China); FAR1 (1:1000; NBP1-89847) from Novus Biologicals (Centennial, CO, USA); GSDMD-NT(1:1000; 10137T), p-MLKL(1:1000; 37333T), MLKL(1:1000; 37705T), p-p65(1:1000; 3033T), and p65(1:1000; 8242T) from Cell Signaling Technology (Danvers, MA, USA); cleaved Caspase-3(1:1000; 68773-1-Ig) and β-actin (1:5000; 20536-1-AP) from Proteintech (Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Control, Fluorescence, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Standard Deviation, Transmission Assay, Electron Microscopy

Journal: Apoptosis

Article Title: Targeting FAR1 to inhibit intestinal macrophage ferroptosis: repurposing irinotecan as a novel sepsis therapy

doi: 10.1007/s10495-026-02419-7

Figure Lengend Snippet: Irinotecan attenuates sepsis-induced ferroptosis through FAR1-dependent regulation. RAW264.7 cells were transfected with a FAR1 overexpression vector (OE-FAR1) or an empty vector (negative control, NC) and treated with LPS/irinotecan as indicated. A , B Western blots and quantitative analysis of NRF2, ACSL4, FAR1, and GPX4 expression ( n = 3 per group). C Representative fluorescence images of intracellular Fe 2+ (FerroOrange; scale bar = 25 μm), lipid peroxides (Liperfluo; scale bar = 25 μm), and ROS (DCFH-DA; scale bar = 25 μm). Flow cytometry analysis of D intracellular Fe 2+ (FerroOrange), E lipid peroxide (Liperfluo), and F ROS levels in RAW264.7 cells 24 h post-treatment. ELISA quantification of G) MDA levels, H the GSH/GSSG ratio, I) TNF-α levels, and J IL-6 levels in cell supernatants ( n = 8 per group). The data are presented as the mean ± standard deviation. ** p < 0.01 versus the Control group + OE-NC; ## p < 0.01 versus the LPS + OE-NC group; && p < 0.01 versus the LPS + OE-NC + Ir group; ns, not significant. Ir, irinotecan; LPS, lipopolysaccharide; OE-NC, overexpression negative control; OE-FAR1, FAR1 overexpression; MDA, malondialdehyde; ROS, reactive oxygen species; GSH/GSSG, glutathione/oxidized glutathione ratio

Article Snippet: Primary antibodies against the following proteins were used: NRF2 (1:1000; A21176), ACSL4 (1:1000; A2041) and GPX4 (1:1000; A11243) from ABclonal (Wuhan, China); FAR1 (1:1000; NBP1-89847) from Novus Biologicals (Centennial, CO, USA); GSDMD-NT(1:1000; 10137T), p-MLKL(1:1000; 37333T), MLKL(1:1000; 37705T), p-p65(1:1000; 3033T), and p65(1:1000; 8242T) from Cell Signaling Technology (Danvers, MA, USA); cleaved Caspase-3(1:1000; 68773-1-Ig) and β-actin (1:5000; 20536-1-AP) from Proteintech (Wuhan, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Negative Control, Western Blot, Expressing, Fluorescence, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Apoptosis

Article Title: Targeting FAR1 to inhibit intestinal macrophage ferroptosis: repurposing irinotecan as a novel sepsis therapy

doi: 10.1007/s10495-026-02419-7

Figure Lengend Snippet: Irinotecan suppresses sepsis-induced ferroptosis through a Gly252-dependent interaction with FAR1. RAW264.7 cells were transfected with a vector encoding the FAR1-G252A mutant and treated with LPS/irinotecan. A Western blots of ACSL4, FAR1, and GPX4 expression. B Representative fluorescence images of intracellular Fe 2+ (FerroOrange; scale bar = 25 μm), lipid peroxides (Liperfluo; scale bar = 25 μm), and ROS (DCFH-DA; scale bar = 25 μm). C quantitative analysis of ACSL4, FAR1, and GPX4 expression ( n = 3 per group). Quantitative analysis of the fluorescence intensity of D intracellular Fe 2+ (FerroOrange), E lipid peroxides (Liperfluo), and F ROS based on the data in panel ( B ). Flow cytometry of G FerroOrange, H Liperfluo, and I ROS levels in RAW264.7 cells 24 h after Control or LPS (500 ng/mL) treatment with or without irinotecan (1 µM) pretreatment. ELISA quantification of J MDA levels, K) the GSH/GSSG ratio, L) TNF-α levels, and M) IL-6 levels in the cellular supernatant ( n = 8 per group). The data are presented as the mean ± standard deviation; ** p < 0.01 versus the Control + FAR1-G252A group; ## p < 0.01 versus the LPS + FAR1-G252A group; ns, not significant. Ir, irinotecan; LPS, lipopolysaccharide; MDA, malondialdehyde; ROS, reactive oxygen species; GSH/GSSG, glutathione/oxidized glutathione ratio

Article Snippet: Primary antibodies against the following proteins were used: NRF2 (1:1000; A21176), ACSL4 (1:1000; A2041) and GPX4 (1:1000; A11243) from ABclonal (Wuhan, China); FAR1 (1:1000; NBP1-89847) from Novus Biologicals (Centennial, CO, USA); GSDMD-NT(1:1000; 10137T), p-MLKL(1:1000; 37333T), MLKL(1:1000; 37705T), p-p65(1:1000; 3033T), and p65(1:1000; 8242T) from Cell Signaling Technology (Danvers, MA, USA); cleaved Caspase-3(1:1000; 68773-1-Ig) and β-actin (1:5000; 20536-1-AP) from Proteintech (Wuhan, China).

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Western Blot, Expressing, Fluorescence, Flow Cytometry, Control, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Apoptosis

Article Title: Targeting FAR1 to inhibit intestinal macrophage ferroptosis: repurposing irinotecan as a novel sepsis therapy

doi: 10.1007/s10495-026-02419-7

Figure Lengend Snippet: Schematic of the Irinotecan–FAR1 (Gly252) axis regulating ferroptotic responses in septic macrophages. During sepsis, upregulated FAR1 promotes a proferroptotic metabolic cascade. The upregulation of FAR1 facilitates fatty acids (FAs)-dependent PUFA-ePL remodeling in conjunction with ACSL4-mediated lipid processing. The accumulation of PUFA-ePL promotes lipid hydroperoxide (LOOH) generation, which may overwhelm GPX4-dependent antioxidant defenses. This metabolic dysregulation increases macrophage ferroptotic susceptibility, contributing to intestinal barrier dysfunction and systemic inflammation. Irinotecan exhibits noncanonical activity through a Gly252-dependent interaction with FAR1. This interaction attenuates FAR1-associated PUFA-ePL metabolic remodeling, thereby limiting LOOH accumulation and preserving GPX4-dependent antioxidant capacity. By attenuating mitochondrial injury associated with metabolic dysregulation and macrophage ferroptotic responses, irinotecan promotes gut barrier restoration and improves survival in preclinical sepsis models. Ir, irinotecan; FA, fatty acid; PUFA-ePLs, polyunsaturated ether phospholipids; LOOH, lipid hydroperoxides

Article Snippet: Primary antibodies against the following proteins were used: NRF2 (1:1000; A21176), ACSL4 (1:1000; A2041) and GPX4 (1:1000; A11243) from ABclonal (Wuhan, China); FAR1 (1:1000; NBP1-89847) from Novus Biologicals (Centennial, CO, USA); GSDMD-NT(1:1000; 10137T), p-MLKL(1:1000; 37333T), MLKL(1:1000; 37705T), p-p65(1:1000; 3033T), and p65(1:1000; 8242T) from Cell Signaling Technology (Danvers, MA, USA); cleaved Caspase-3(1:1000; 68773-1-Ig) and β-actin (1:5000; 20536-1-AP) from Proteintech (Wuhan, China).

Techniques: Activity Assay, Preserving

Journal: Developmental cell

Article Title: Phosphorylated Lamin A/C in the nuclear interior binds active enhancers associated with abnormal transcription in progeria

doi: 10.1016/j.devcel.2020.02.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: We cloned Lamin A or Lamin C cDNAs with S22 and S392 mutations or without mutations into the allin-one doxycycline inducible lentivirus vector pCW57-MCS1-P2A-MCS2-PGK-Blast (gift from Adam Karpf; Addgene plasmid #80921) ( Barger et al., 2019 ) using HiFi assembly (New England Biolabs E2621).

Techniques: Recombinant, Binding Assay, Sequencing, Next-Generation Sequencing, Cloning, Plasmid Preparation, Software