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: 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