cd36 knockout mice  (Jackson Laboratory)

Journal: iScience

Article Title: Angiogenesis-independent VEGF signaling enhances exercise capacity by increasing fat oxidation in mice fed sulfur amino acid-restricted diets

doi: 10.1016/j.isci.2025.114148

Figure Lengend Snippet: Transcriptomics across muscle depots reveal metabolic shift from glycolytic toward oxidative (A) Experimental set up and color scheme used throughout and . (B) Fold changes of transcripts associated with fatty acid (FA) catabolism and transport as identified in supplementary Extended Data A in muscle of male mice ( n = 6) given ad libitum access to sulfur amino acid-restricted (SAAR) versus control (Con) diet for seven days. (C) Pathway enrichment analysis of genes showing significant diet-by-muscle interaction effects. (D) Fold changes (SAAR vs. Con) of TCA cycle genes in EDL and soleus. (E) Representative blots of electron transport chain complexes. (F) Quantification of relative protein abundance normalized to vinculin of SDHB for EDL and soleus ( n = 6) of male mice given ad libitum access to SAAR versus Con diet on day seven. (G) Representative blots of CD36, LPL, and PDK4 and vinculin ( top ) and quantification of relative protein abundance normalized to vinculin of CD36, PDK4, and LPL ( bottom ) in EDL ( n = 6) of male mice given ad libitum access to SAAR versus Con diet on day seven. (H) Representative blots of CD36, LPL, and PDK4 and vinculin ( top ) and quantification of relative protein abundance normalized to vinculin of CD36, PDK4, and LPL from blots ( bottom ) in soleus ( n = 6) of male mice given ad libitum access to SAAR versus Con diet on day seven. All panels represent data from mice that were not subjected to endurance running. All data are shown as mean, and error bars indicate SD unless otherwise noted; p values indicate the significance of the difference by Student t test between diets; significance is determined by p < 0.05. See also and . Raw data files for E are available in the supplement as ZIP file.

Article Snippet: Wild type (WT) C57BL/6J and Cd36 LoxP / LoxP mice ( Cd36 tm1.1Ijg /J) were purchased from Charles River (Freiburg im Breisgau, Germany).

Techniques: Control, Quantitative Proteomics

Journal: iScience

Article Title: Angiogenesis-independent VEGF signaling enhances exercise capacity by increasing fat oxidation in mice fed sulfur amino acid-restricted diets

doi: 10.1016/j.isci.2025.114148

Figure Lengend Snippet: SAAR increases muscle lipid flux without altering lipid pool sizes (A) Experimental design and color scheme used in A–3E and A. (B) Circulatory carbon flux ( n = 10–13) of 13 C 18 -U-Linolate of jugular-vein-catheterized male mice given ad libitum access to sulfur amino acid-restricted (SAAR) versus control (Con) diet for seven days. (C) Ex vivo β-oxidation measured by incorporation of 3 H-palmitic acid in 3 H-H 2 O in muscles of male mice fed a Con or SAAR diet for seven days ( n = 15). (D) Representative fluorescence images ( left ) of BODIPY 493:503 (green), WGA647 (red), and DAPI (blue) staining in EDL cross sections (scale bar, 50 μm) and quantification of Bodipy + Intensity within fibers ( right ) of male mice fed a Con or SAAR for seven days ( n = 6). (E) Lipidomics analysis from muscle of male mice fed a Con or SAAR diet for seven days ( n = 6), summarized as normalized ion counts of each main lipid class. (F) Experimental set up and color scheme used in G, 3H, and D–S3I. (G) Percent change in body weight ( n = 16) of male WT and EC CD36−/− mice given ad libitum access to SAAR versus Con diet after seven days. (H) Distance ran during a one-time maximal endurance test (n = 9–15) of male WT and EC CD36−/− mice given ad libitum access to SAAR versus Con diet on day seven. B–E represent data from mice that were not subjected to endurance running, G and H represent data from mice subjected to maximal endurance testing. All data are shown as mean, and error bars indicate SD unless otherwise noted; p values indicate the significance of the difference by Student t test between diets, or two-way ANOVA with Sidak multiple comparisons test between diets and muscle or genotype; significance is determined by p < 0.05. See also and , , and .

Article Snippet: Wild type (WT) C57BL/6J and Cd36 LoxP / LoxP mice ( Cd36 tm1.1Ijg /J) were purchased from Charles River (Freiburg im Breisgau, Germany).

Techniques: Control, Ex Vivo, Muscles, Fluorescence, Staining

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Schematic workflow of the cardiac tissues single-cell sequencing. Cells were isolated by mechanical and enzymatic dissociation from the ventricular tissues of adult mice. Live cells were used for scRNA-seq using the 10X Genomics platform. b Uniform Manifold Approximation and Projection (UMAP) plot of the aggregate of all sequenced cardiac cells identified 9 broad cell types after unsupervised clustering. Violin plot showing the expression levels of cardiomyocyte genes Acta2 and Atp2a2 signature in all identified cell types. c Volcano plot of differentially expressed genes in MI vs. Sham cardiomyocytes (fold change > 1.2, P -value < 0.05). P -values were assessed by the Wilcoxon rank-sum test. d A Venn diagram showing the overlap of detected cardiac DEPs and DEGs of cardiomyocytes. e GO pathway enrichment and ( f ) KEGG pathway enrichment in the overlapping data (Significance was calculated by a hypergeometric test). g Gene expression visualized in UMAP plots for CD36, Fabp4 and Fabp5 in cardiomyocytes of Sham and MI mice.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Sequencing, Isolation, Expressing, Gene Expression

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Western-blot analysis of CD36 in different fractions of myocardium. n = 9 for total CD36; n = 5 for plasma membrane CD36; n = 8 for subcellular CD36. b CD36 mRNA level in isolated adult cardiomyocytes from Sham and MI mice. n = 6. c Western-blot analysis of CD36 in different fractions of isolated adult cardiomyocytes from Sham and MI mice. n = 6 for total CD36 and plasma membrane CD36; n = 8 for subcellular CD36. d Immunostaining assay was used to analyze the colocalization of CD36 (green) and Na + /K + -ATPase (red) in isolated adult cardiomyocytes. Trace outline is used for line-scan analysis of relative fluorescence intensities of CD36 with Na + /K + -ATPase signals. Scale bar = 100 µm. Data are presented as means ± SEM. Statistical significance was assessed by a two-tailed unpaired Student t test ( a – c ). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Western Blot, Clinical Proteomics, Membrane, Isolation, Immunostaining, Fluorescence, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Schematic diagram of the study design, CD36-specific inhibitor Sulfosuccinimidyl oleate sodium (SSO) or solvent once a week intraperitoneally for 4 weeks after MI. Echocardiography was performed at 4 weeks post-MI. b Ejection fraction (EF) and fractional shortening (FS) values were determined to evaluate cardiac function by echocardiography in mice of each group. n = 8 per group. c Averaged infarct size by 2,3,5-triphenyltetrazolium chloride (TTC) staining. n = 5. d , e Histological analysis of myocardial tissue sections with Masson staining ( d ) and hematoxylin-eosin (H&E) staining ( e ). n = 6 per group. f Representative transmission electron microscopy images of myocardium from mice with or without SSO treatment after MI. n = 5 per group. g , h Cardiac ceramide ( g ) and DAG levels ( h ) after treatment with SSO detected by ELISA assay. n = 8 for ceramide, n = 7 for DAG. i , j Quantitation analysis of Cardiac tissue malondialdehyde (MDA) ( i ) and superoxide dismutase (SOD) ( j ) levels. n = 9 per group. Data are presented as means ± SEM. Statistical significance was assessed by Kruskal-Wallis, followed by the false discovery rate [FDR] method of Benjamini and Hochberg test ( b , i ) and One-way ANOVA, followed by Tukey post hoc multicomparisons test ( c , d , g , h and j ). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Solvent, Staining, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Quantitation Assay

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Protein levels of palmitoylated CD36 in hearts of Sham and MI mice. n = 6 per group. b Palmitoylated CD36 protein levels in PA (160 µM, 24 h) or 2 bp (25 µM, 24 h) treated cardiomyocytes (NMVCs). n = 9 per group. c Western blotting analysis of plasma membrane and total CD36 protein in PA or 2 bp treated cardiomyocytes (NMVCs). n = 6 per group. d , e Bivariate flow cytometry analysis was conducted to investigate the impact of CD36 palmitoylation on FA uptaking in cardiomyocytes (NMVCs). d Mean Fluorescence Intensity of Bodipy FL-C16. n = 6. e The values provided are in arbitrary fluorescence units. In cases where there were 3 cells or fewer for a given PE fluorescence channel, the point is plotted at 0 on the y -axis. The blue dots plotted along the x -axis indicate significant differences in FA uptaking values between Control and PA or 2 bp treatment cells with similar CD36 expression levels (determined by Student’s t-tests reaching the 99.9% confidence value) . f – h Palmitoylated CD36 protein and plasma membrane CD36 protein levels in PA or 2 bp treated cardiomyocytes (NMVCs) after 24 h of hypoxia induction. n = 5 for palmitoylated CD36; n = 6 for surface CD36. i , j FA uptake ( i ) and binding ( j ) analyses were conducted on cardiomyocytes (NMVCs) treated with PA or 2 bp following 24 h of hypoxia induction. n = 7 for cell uptake; n = 5 for cell binding. k , l Ceramide ( k ) and DAG ( l ) levels in cardiomyocytes (NMVCs) in each group. n = 8 for ceramide; n = 7 for DAG. Data are presented as means ± SEM. Statistical significance was assessed by a two-tailed unpaired Student t test ( a ), One-way ANOVA, followed by Tukey post hoc multicomparisons test ( b , c [Surface CD36], d and f – l ) and Kruskal-Wallis, followed by false discovery rate [FDR] method of Benjamini and Hochberg test ( c [Total CD36]). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Western Blot, Clinical Proteomics, Membrane, Flow Cytometry, Fluorescence, Control, Expressing, Binding Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Total CD36 protein expression in cardiomyocyte (NMVCs) was detected by FACS. n = 6 per group. b The plasma membrane and cytoplasm fraction were isolated from cardiomyocytes (NMVCs), and the relative distribution of CD36 protein in different fractions was analyzed by western blot. n = 6 for surface CD36; n = 10 for subcellular CD36. c , d ( c ) The dual-parameter contour plot correlating FL-C16 uptaking against CD36 expression, as well as a histogram showing the Mean Fluorescence Intensity (MFI) for each group. n = 7. d Cysteine mutation in the palmitoylation site of CD36 reduced FA transport. The blue points plotted along the x-axis indicate significant differences in FA uptaking values between WT-CD36 and AA-SS-CD36 cells with similar CD36 expression levels (determined by Student’s t tests reaching the 99.9% confidence value). e , f Ceramide ( e ) and DAG ( f ) levels in cardiomyocytes (NMVCs) transfected with WT-CD36 and AA-SS-CD36 after 24 h of hypoxia induction. n = 5 for ceramide; n = 7 for DAG. g The mitochondrial oxygen consumption rate (OCR) in cardiomyocytes (NMVCs) transfected with WT-CD36 and AA-SS-CD36 after 24 h of hypoxia induction. n = 5 per group. h Mean fluorescence intensity of MitoSOX in cardiomyocytes (NMVCs) transfected with WT-CD36 and AA-SS-CD36 after 24 h of hypoxia induction. n = 6 per group. Data are presented as means ± SEM. Statistical significance was assessed by One-way ANOVA, followed by Tukey post hoc multicomparisons test ( a and e – h ), two-tailed Mann-Whitney U test ( b [surface CD36], c ) and two-tailed unpaired t test ( b [subcellular CD36]). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Expressing, Clinical Proteomics, Membrane, Isolation, Western Blot, Fluorescence, Mutagenesis, Transfection, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Schematic diagram of the study design, CD36 knockout (CD36KO) mice were injected with adenoassociated virus 9 (AAV9)–cTnT (cardiac troponin T2)-WT-CD36 (wildtype CD36) or AAV9-cTNT-AA-SS-CD36 (CD36 with mutated palmitoylation sites) via the tail vein. After 4 weeks, the mice underwent left anterior descending coronary artery ligation surgery, followed by echocardiographic measurements on cardiac function. b Protein levels of CD36 in total homogenates and different fractions of WT-CD36 and AA-SS-CD36 mice. n = 6 for total CD36; n = 4 for surface membrane CD36.; n = 8 for subcellular CD36. c – e Echocardiography determined EF and FS values to assess cardiac function after sham or LAD surgery in WT-CD36 and AA-SS-CD36 mice. n = 7 per group. f – h Histological sections with hematoxylin-eosin (H&E) staining and Masson staining of myocardial tissues from WT-CD36 and AA-SS-CD36 mice. n = 5 per group. i Representative transmission electron microscopy images of cardiac tissues from WT-CD36 and AA-SS-CD36 mice (yellow arrow indicates autophagosome). n = 5 per group. j – m Changes in cardiac biomarker levels: cardiac ceramide ( j ), DAG ( k ), malondialdehyde (MDA) ( l ) and superoxide dismutase (SOD) ( m ) levels in WT-CD36 and AA-SS-CD36 mice. n = 10 for ceramide; n = 8 for DAG; n = 9 for MDA and SOD. Data are presented as means ± SEM. Statistical significance was assessed by One-way ANOVA, followed by Tukey post hoc multicomparisons test ( b – g and k ) and Kruskal-Wallis, followed by the false discovery rate [FDR] method of Benjamini and Hochberg test ( j , l and m ). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Knock-Out, Injection, Virus, Ligation, Membrane, Staining, Transmission Assay, Electron Microscopy, Biomarker Discovery

Journal: Nature Communications

Article Title: Inhibiting CD36 palmitoylation improves cardiac function post-infarction by regulating lipid metabolic homeostasis and autophagy

doi: 10.1038/s41467-025-61875-y

Figure Lengend Snippet: a Western blotting analysis of LC3II and SQSTM1 protein levels in myocardium from WT-CD36 and AA-SS-CD36 mice. n = 7. b Western blotting analysis of LC3II and SQSTM1 protein levels in cardiomyocytes transfected with WT-CD36 or AA-SS-CD36 for 48 h. n = 10. c Overlap between CD36 binding proteins, as assessed by CoIP-MS analysis and proteins in the Autophagy Database ( http://autophagy.info/ ). d Endogenous CD36 immunoprecipitation followed by anti-CD36 and anti-PGAM5 western blot analysis of cardiomyocytes (NMVCs). n = 4. e Quantification of Cyto-ID staining in cardiomyocytes (NMVCs) transfected with WT-CD36 or AA-SS-CD36 for 48 h, and then treated with 500 nM rapamycin (Rapa) and 60 µM CQ, the combination for 16 h. n = 6 per group. f – h Western blotting analysis of the mitochondria expression of CD36 and PGAM5 in cardiomyocytes (NMVCs) transfected with WT-CD36 or AA-SS-CD36. n = 7 per group. i Relative PGAM5 mRNA levels in cardiomyocytes transfected with WT-CD36 or AA-SS-CD36 for 48 h. n = 5. j Time course of PGAM5 protein stability in cardiomyocytes (NMVCs) transfected with WT-CD36 or AA-SS-CD36 after treatment with CHX (10 µg/ml). n = 6. k Ubiquitination of exogenous PGAM5 in cardiomyocytes (NMVCs) transfected with WT-CD36 or AA-SS-CD36. Data are representative of six independent experiments. l The mitochondria expression of phosphorylation Fundc1 and total Fundc1 protein in cardiomyocytes (NMVCs) transfected with WT-CD36 or AA-SS-CD36. n = 6 for phosphorylation Fundc1; n = 7 for total Fundc1. Data are presented as means ± SEM. Statistical significance was assessed by Kruskal-Wallis, followed by false discovery rate [FDR] method of Benjamini and Hochberg test ( a ), two-tailed unpaired Student t test ( b , d , g , i and l ), two-way ANOVA, followed by Tukey post hoc multicomparisons test ( e , j ) and two-tailed Mann-Whitney U test ( h ). Source data are provided as a file.

Article Snippet: CD36 knockout (KO) mice were generated using the CRISPR/Cas9 technique by Cyagen Biosciences Co., Ltd (Cyagen, Jiangsu, China).

Techniques: Western Blot, Transfection, Binding Assay, Immunoprecipitation, Staining, Expressing, Ubiquitin Proteomics, Phospho-proteomics, Two Tailed Test, MANN-WHITNEY