goat polyclonal  (R&D Systems)

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: Mice that lack CD36-expressing B cells exhibit compromised autoimmunity. A The immunization strategy employed for the Mb1 cre and Cd36 fl/fl Mb1 cre mice. Mice were immunized four times weekly with apoptotic cells (4xAC), while PBS was used as the negative control. B The gating strategy and frequency of germinal center (GC) B cells were examined. C Measurement of anti-DNA IgG levels via ELISA. The data are representative of three independent experiments. * P < 0.05, ** P < 0.01 and *** P < 0.001 (Mann–Whitney)

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques: Expressing, Negative Control, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: CD36-associated proteins were identified through co-immunoprecipitation followed by mass spectrometry analysis. A The pseudocolor plot illustrates the staining of CD36 in B cells that were either stimulated with CPG (CPG Stim) or unstimulated (Uns). B Confocal microscopy was used to analyze CD36 staining in B cells treated with CPG. C Immunoblot analysis of soluble proteins derived from B cells stimulated with CPG was subsequently performed using an anti-CD36 antibody. The total B cell protein served as the loading (positive) control referred to as Input (Inp). The B cell protein was immunoprecipitated using isotype (negative) control IgG antibodies (IgG) or anti-CD36 antibodies (CD36). D Evidence of the CD36 protein network predicted by the STRING, demonstrating its functional association with the top ten proteins. Network nodes represent proteins. Colored nodes represent the query protein and its first shell of interactors, while white nodes represent the query protein and the second shell of interactors. Filled nodes or empty nodes indicate predicted or unpredicted 3D structures, and edges represent protein–protein associations. Line color indicates the different types of interaction evidence. E Confidence view of the CD36 protein network. The line of thickness visually depicts stronger associations. F , G Gene Ontology enrichment analysis was also conducted to investigate the biological processes and molecular functions associated with the proteins identified through anti-CD36 immunoprecipitation. The data are representative of three independent experiments. * P < 0.05, ** P < 0.01 and *** P < 0.001 (Mann–Whitney)

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques: Immunoprecipitation, Mass Spectrometry, Staining, Confocal Microscopy, Western Blot, Derivative Assay, Positive Control, Negative Control, Functional Assay, MANN-WHITNEY

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: The protein interaction between CD36 and FcγRIIb was confirmed through various experimental approaches. A CD36 and FcγRIIb staining analysis of CPG-stimulated B cells using confocal microscopy. B The proteins immunoprecipitated by anti-CD36 antibodies were detected using anti-FcγRIIb antibodies in B cells stimulated with cytidine phosphate guanosine (CPG) or lipopolysaccharide (LPS). The total B cell protein served as the loading (positive) control referred to as Input (Inp). The B cell protein was immunoprecipitated using isotype (negative) control IgG antibodies (IgG) or anti-CD36 antibodies (CD36). C , D Docking scores and ligand mean square deviation (rmsd) for the models of CD36 docking with FcγRIIb by HDOCK SERVER. The X-axis represents the rank of the top 10 models, and the Y-axis shows the docking energy scores in C . The ligand rmsd is calculated by comparing the ligands in the CD36 model with the modeled FcγRIIb structures in D . E , F The front and back sides of the model 1 of CD36 (multicolor) docking with FcγRIIb (purple)

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques: Staining, Confocal Microscopy, Immunoprecipitation, Positive Control, Negative Control

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: Complex template information for CD36 and FcγRIIb

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques:

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: Interface residue pairs and ligand root mean square deviation (rmsd) (Å) for Model 1 of CD36 docking with FcγRIIb

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques: Residue

Journal: Cellular & Molecular Biology Letters

Article Title: Unveiling the hidden role of the interaction between CD36 and FcγRIIb: implications for autoimmune disorders

doi: 10.1186/s11658-024-00593-7

Figure Lengend Snippet: Loss of FcγRIIb in mice downregulates CD36 expression in B cells. A The gating strategy and scatter plot depict CD36 staining in marginal zone B cells (MZB) (CD19 + CD21 + CD23 mid ) from both wild-type (WT) and Fcgr2b knockout ( Fcgr2b KO) mice determined via flow cytometry. B The immunization strategy involved apoptotic cells (4xAC) in WT and Fcgr2b KO mice. PBS served as the negative control. C The CD36 staining was performed on MZB in WT and Fcgr2b KO mice under injections. D , E The gating strategy and scatter plot illustrate the CD36 staining pattern in germinal center B cells (GC) (B220 + IgD − CD95 + GL-7 + ) of and plasma cells (PC) (B220 int CD138 + ) in immunized WT and Fcgr2b KO mice. F The FcγRIIb staining on B cells, follicular B cells (FOB), and marginal zone B cells (MZB). G The immunization strategy was used for both WT and Cd36 knockout ( Cd36 KO) mice with four times weekly apoptotic cells (4xAC) or PBS. H The FcγRIIb staining of MZB in WT and Cd36 KO mice after treatments. The data are representative of three independent experiments. * P < 0.05, ** P < 0.01 and *** P < 0.001 (Mann–Whitney)

Article Snippet: To conduct immunoprecipitation, a total of 2 µg of goat anti-mouse CD36 antibody (R&D) or normal goat IgG (Millipore) was incubated with Pierce™ Protein A/G Magnetic Beads (Thermo Scientific™) overnight.

Techniques: Expressing, Staining, Knock-Out, Flow Cytometry, Negative Control, MANN-WHITNEY

Journal: Plants

Article Title: Botanical Antcin K Alleviates High-Fat Damage in Palm Acid Oil-Treated Vascular Endothelial Cells and Macrophages

doi: 10.3390/plants11212812

Figure Lengend Snippet: Antcin K treatments effectively reduced the expression of CD36 in palm oil-treated vascular endothelial cells. ( A ) Immunofluorescence staining to examine expressions of CD36 (green) and nuclei (blue, DAPI) in vascular endothelial cells with and without palm acid oil (PA) treatments, and with and without antcin K treatments. ( B ) Comparison of the quantified expression of CD36 of vascular endothelial cells with and without 0.75 mM palm acid oil (PA) treatment, and with and without 20 μg/mL antcin K treatment ( n = 3 for each group; values are presented as mean ± SEM, * p < 0.05, ** p < 0.01, one-way ANOVA followed by the Student–Newman–Keuls multiple comparison post hoc test).

Article Snippet: After washing three times with PBS, the tissues or cells were added 2% BSA to dilute the primary antibody of human anti-mouse VCAM-1 (1:200; sc-13160; Santa Cruz Biotechnology Inc., Dallas, TX, USA), rabbit anti-mouse KLF4 polyclone antibody (membrane protein) (1:200; GTX101508; Genetex, Irvine, CA, USA), or goat anti-mouse CD36 monoclonal antibody (1:200; sc-7309; Santa Cruz Biotechnology Inc.), and then incubated overnight in a humidified dark box at 4 °C.

Techniques: Expressing, Immunofluorescence, Staining, Comparison

Journal: Plants

Article Title: Botanical Antcin K Alleviates High-Fat Damage in Palm Acid Oil-Treated Vascular Endothelial Cells and Macrophages

doi: 10.3390/plants11212812

Figure Lengend Snippet: Possible therapeutic mechanisms of antcin K in alleviating the high-fat damage of vascular endothelial cells and macrophages in blood vessels. Antcin K treatment can (1) decrease the expression of VCAM-1 that alleviates circulating monocytes adhering to endothelial cells and migrating into subendothelial space; (2) reduce ROS generation and oxidative stress that alleviate the oxidative modification of lipoproteins and phospholipids; (3) enhance the expression of KLF4 in macrophages and endothelial cells that decrease lipid deposition and prevent macrophages converting into foam cells; (4) decrease the content of TNF-α and IL-1β; and (4) decrease the expression of CD36 that slows transformation of macrophages and endothelial cells to foam cells.

Article Snippet: After washing three times with PBS, the tissues or cells were added 2% BSA to dilute the primary antibody of human anti-mouse VCAM-1 (1:200; sc-13160; Santa Cruz Biotechnology Inc., Dallas, TX, USA), rabbit anti-mouse KLF4 polyclone antibody (membrane protein) (1:200; GTX101508; Genetex, Irvine, CA, USA), or goat anti-mouse CD36 monoclonal antibody (1:200; sc-7309; Santa Cruz Biotechnology Inc.), and then incubated overnight in a humidified dark box at 4 °C.

Techniques: Expressing, Modification, Transformation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

doi: 10.3389/fcell.2021.749153

Figure Lengend Snippet: HDL-uptake by Plasmodium -infected erythrocytes. (A) Uptake of HDL components by P. b erghei-pRBCs. Phospholipid and ester components were concomitantly observed around parasite, but HDL was not taken up by the nRBCs (arrows). Scale bar: 5 μm. (B) Pb -pRBCs western blotted for Apo A-I, using a polyclonal primary Ab reactive to both mouse and human Apo A-I. The membrane fraction showed Apo A-I positivity with or without the addition of human HDL. (C) Flow cytometric analysis of DiO-labelled HDL uptake. Synchronized ring-stage P. falciparum incubated for 24 h with 500 μg/ml of HDL. HDL uptake was competitively inhibited by unlabeled HDL. (D) Chemical structure of BLT-1. (E) BLT-1 dose-dependent inhibition of DiO-labelled HDL uptake to Pb -pRBCs. The numbers in panels represent fraction (%) of DiO-positive cells out of total cells.

Article Snippet: Exosomes from 50 μl of equally pooled plasma from five mice per group were sequentially incubated at room temperature for 1 h with goat polyclonal anti-CD36 Ab (AF2519, R and D Systems) and HRP-labelled bovine anti-goat IgG heavy and light chains, known to be minimally cross-reactive with mouse and rat serum proteins (805–035-180, Jackson ImmunoResearch, West Grove, PA, United States).

Techniques: Infection, Western Blot, Membrane, Incubation, Inhibition

Journal: Frontiers in Cell and Developmental Biology

Article Title: Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

doi: 10.3389/fcell.2021.749153

Figure Lengend Snippet: Immunocytochemical analysis of Pb -pRBCs for scavenger receptors and CD41. (A) Immunocytochemical analysis of Pb -pRBCs. The CD36 signals (pink) were weak in the pRBCs on days 5 and 8 (ring stage), but were abundantly detected on days 11 and 14 (schizonts). On day 11, CD36 appeared as a spot corresponding to each nucleus in the schizont’s body. SR-B1, which was detected in the pRBC membrane, was negligible in the internal parasites (arrow). Scale bar: 5 μm (B) Immunocytochemical analysis of CD36 in BM-transplanted mouse erythrocytes 6 days after Pb infection. CD36 (pink) was detected in pRBCs from the Kusabira-Orange mouse, but not in the BM-transplanted or CD36 −/− mouse. Scale bar: 5 μm. (C) Immunocytochemical analysis of pRBCs. CD41 (pink) signals were absent on days 5 and 8 (ring stage), but a small signal emanated from the internal parasites on day 8. CD41-specific fluorescence became abundant on days 11 and 14 (schizont stage). On day 11, CD41 appeared as spots corresponding to nuclei in the schizont’s body, as was also seen with CD36. Scale bar: 5 μm. (D) Co-localization of CD36 (red) and CD41 (green). Scale bar: 5 μm.

Article Snippet: Exosomes from 50 μl of equally pooled plasma from five mice per group were sequentially incubated at room temperature for 1 h with goat polyclonal anti-CD36 Ab (AF2519, R and D Systems) and HRP-labelled bovine anti-goat IgG heavy and light chains, known to be minimally cross-reactive with mouse and rat serum proteins (805–035-180, Jackson ImmunoResearch, West Grove, PA, United States).

Techniques: Membrane, Infection, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

doi: 10.3389/fcell.2021.749153

Figure Lengend Snippet: Analyses of the exosomes isolated from Pb -parasitized mouse plasma or from cultured cells differentiated from CMK11-5 cells. (A) Electron micrograph of the plasma exosomes (arrows) on day 8 post-infection. Scale bar: 333 nm. (B) Western blot of exosomes from plasma, platelets, and pRBCs. CD36, CD41 and an exosome marker (ALIX) were detected on days 5 and 8 post-infection. (C) Detection of exosomes doubly-positive for CD36 and CD41 by sandwich ELISA (duplicated). Each group of exosomes was isolated from the ‘pooled plasma’ of five mice. PBS and exosomes from a CD36 −/− mouse were used as negative controls. On days 5 and 8 post-infection, increased OD450 nm was observed. (D) immunoblot analysis for CD36 and CD41 in cultured cells differentiated from CMK11-5 cells or the exosomes released from those cells. (E) Immunocytochemical analysis of erythrocytes from a Pb -parasitized CD36 −/− mouse 6 h after adding CMK11-5-derived exosomes. CD36 and CD41 were detected from internal Pb (pink). NC: parasitized erythrocytes incubated with CMK11-5-derived exosomes and immunostained without primary Abs but with secondary Abs. Scale bar: 5 μm. (F) Trafficking of DiI-labelled exosomes derived from CMK11-5 cells into erythrocytes 1 h after exosome addition. DiI was observed in pRBCs (red) but not in uninfected ones. Scale bar: 5 μm.

Article Snippet: Exosomes from 50 μl of equally pooled plasma from five mice per group were sequentially incubated at room temperature for 1 h with goat polyclonal anti-CD36 Ab (AF2519, R and D Systems) and HRP-labelled bovine anti-goat IgG heavy and light chains, known to be minimally cross-reactive with mouse and rat serum proteins (805–035-180, Jackson ImmunoResearch, West Grove, PA, United States).

Techniques: Isolation, Clinical Proteomics, Cell Culture, Infection, Western Blot, Marker, Sandwich ELISA, Derivative Assay, Incubation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

doi: 10.3389/fcell.2021.749153

Figure Lengend Snippet: In vivo Pb survival and growth (parasitemia). (A) Kaplan-Meier survival curve of Pb -infected mice. CD36 −/− mice survived for significantly longer than the C57BL6 mice. (B) Parasitemia during Pb infection.

Article Snippet: Exosomes from 50 μl of equally pooled plasma from five mice per group were sequentially incubated at room temperature for 1 h with goat polyclonal anti-CD36 Ab (AF2519, R and D Systems) and HRP-labelled bovine anti-goat IgG heavy and light chains, known to be minimally cross-reactive with mouse and rat serum proteins (805–035-180, Jackson ImmunoResearch, West Grove, PA, United States).

Techniques: In Vivo, Infection

Journal: Frontiers in Cell and Developmental Biology

Article Title: Malaria Parasites Hijack Host Receptors From Exosomes to Capture Lipoproteins

doi: 10.3389/fcell.2021.749153

Figure Lengend Snippet: Proposed mechanism of exosome-based CD36 delivery and HDL capture in pRBCs. Pf EMP1 in P. falciparum and proteins that have similar binding ability to CD36 in mice malaria catch exosomes to hijack the receptors.

Article Snippet: Exosomes from 50 μl of equally pooled plasma from five mice per group were sequentially incubated at room temperature for 1 h with goat polyclonal anti-CD36 Ab (AF2519, R and D Systems) and HRP-labelled bovine anti-goat IgG heavy and light chains, known to be minimally cross-reactive with mouse and rat serum proteins (805–035-180, Jackson ImmunoResearch, West Grove, PA, United States).

Techniques: Binding Assay

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Representative histology and immunohistochemistry of a stomach from a wildtype (WT) mouse (left) and a Cd36 −/− mouse (right) showing the altered organization of acid-producing parietal cells (PC) around blood vessels in gastric glands (blue asterisks). b CD36 expression in the corpus is most abundant on endothelial cells and is also detected in PCs. c In PCs, CD36 (red) is at the basolateral membrane (dotted yellow line) and is excluded from the apical membrane marked by ezrin (magenta) and in contact with the lumen (blue dotted line). d , e Immunostaining for ghrelin and gastrin (right insets: magnification of cell markers colocalized with CD36). Scale bar: 50 µm, except for c and inserts in d , e (10 µm). b – e are stomachs from WT mice. Scale bar: 50 µm. f – i mRNA for hormones in fasted or 4 h refed mice. All mRNA adjusted to 36b4. j Plasma gastrin and k plasma leptin in fasted or 4 h refed mice. SST somatostatin. Significance was calculated using a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001. Data are means ± SEM, f – i : n = 5, j n = 7, k n = 8.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Immunohistochemistry, Expressing, Membrane, Immunostaining, Clinical Proteomics

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a – c Impaired response of plasma leptin, insulin, and pancreatic polypeptide in fasted mice and 15 min after an oral high fat meal. d Stomach acetylcholine levels in 4 h refed mice. e, f Gastric secretions at 1 h after pylorus ligation in 18 h fasted mice given i.p. saline (Control), carbachol (CCh, 60 µg/kg) or histamine (H, 10 mg/kg). e Gastric juice pH. f Immunoblots of gastric juice; quantification of gastric intrinsic factor (GIF)/total protein in juice showing reduced GIF output in Cd36 −/− mice given carbachol. g Chief cells markers by qPCR and immunostaining (green) in corpus of wildtype (WT) and Cd36 −/− mice, scale: 50 µm. h mRNA of muscarinic receptor 1 (M1r) in the corpus of WT and Cd36 −/− mice. Significance was calculated using a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons or a two-sided unpaired t test ( d and h ). * P < 0.05, ** P < 0.01. a – c n = 4–5 per condition/group; d : n = 7–12; e, f n = 3–4; g, h n = 5.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Clinical Proteomics, Ligation, Saline, Control, Western Blot, Immunostaining

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Transmission electron microscopy (TEM) of parietal cells in the Cd36 −/− corpus show a lack of mitochondria-free area at the basolateral side (blue asterisks) and ECM deposits between the PC membrane and blood vessels (white arrows). Scale bars: 6 µm (top) and 1 µm (bottom) panels. b mRNA of ECM proteins fibronectin, laminin and collagen1α in nonfasted mice. c Immunostaining of fibronectin (green) in corpus of WT and Cd36 −/− mice. Scale bar: 50 µm. d mRNA of inflammation and neutrophil markers. Significance was calculated using a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001. Data are means ± SEM. b n = 7, d n = 5.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Transmission Assay, Electron Microscopy, Membrane, Immunostaining

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a–c Representative histology and immunohistochemistry of wildtype (WT) and Cd36 −/− mice that were either untreated (control), or injected with TAM for 3 days followed by 5 days of recovery (TAM + D5); a proliferation marker ki67; b vascular endothelial growth factor B (VEGFB), red; ezrin, green; c GIF, green; GSII, red. Scale bar: 50 µm. Quantification of d ki67 + cells and e parietal cells per gastric unit, f gland length, and g GIF/GSII + cells per gastric unit (yellow). Significance was calculated using a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons. * P < 0.05, ** P < 0.01. Data are means ± SEM, n = 3 mice/group.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Immunohistochemistry, Control, Injection, Marker

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Tissue uptake of oleic acid in wildtype (WT) and Cd36 −/− mice. Four-month-old mice 16 h fasted mice, were given a retro-orbital injection of [ 3 H]oleic acid and tissues collected 5 min later for measuring radioactivity. b High-resolution respirometry (Oxygraph-2k) of permeabilized corpus mucosa from WT and Cd36 −/− mice. Leak: plus malate (1 mM), glutamate (10 mM) and pyruvate (5 mM). OxPHOS: Oxygen consumption, plus ADP (5 mM) and succinate (10 mM). ETS: Maximum flux after FCCP uncoupling. c Ratio of OxPHOS/leak in WT and Cd36 −/− mucosa showing trend for lower coupling efficiency. d Cd36 −/− stomachs have lower levels of short-chain acylcarnitines but accumulate long-chain acylcarnitines. e, f cardiolipin (CL) enrichment with ω6 PUFA. g transmission electron microscopy (TEM) showing altered morphology of PC mitochondria (arrow) in Cd36 −/− mice, scale bar: 2 µm. h Mitochondrial circularity and aspect ratio (major/minor axis). Significance was calculated using a two-sided unpaired t test or a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons ( e , f , and h ). * P < 0.05, * P < 0.01, *** P < 0.001, **** P < 0.0001. Data are means + SEM. a n = 4–5; b– c n = 4; d– f n = 7–12; h n = 600 mitochondria from 9 PCs/genotype.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Injection, Radioactivity, Transmission Assay, Electron Microscopy

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Heat map of top 60 metabolites/lipids in fasted and re-fed wildtype (WT) and Cd36 −/− stomach. b The sum of identified diacylglycerol (DAG) and triacyglycerol (TAG) species was reduced in Cd36 −/− stomachs. c Cd36 deficient stomachs accumulate more phospholipids with d higher % of 20:4. e Cd36 −/− mice have lower mRNA levels of TAG synthesis enzyme glycerol-3-phosphate acyltransferase 4 (GPAT4) but increased levels of the TAG lipolytic enzyme adipose triglyceride lipase (ATGL). f Schematic summary of key steps in TAG synthesis with intermediates that serve as precursors of other lipid species, such as phospholipids (PL). CL cardiolipin, FA fatty acids, MAG monoacylglycerol, PC phosphatidylcholine, PE phosphatidylethanolamine, PG phosphatidylglycerol, PI, phosphatidylinositol, PS, phosphatidylserine, PUFA, polyunsaturated FA. Significance was calculated using a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data are means ± SEM, n = 7–12 mice/group for a – d , and n = 5 for e .

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques:

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Gastric sections from floxed controls (left panels) and EC- Cd36 -/- mice (right panels) showing absence of CD36 expression in the gastric endothelium and CD36 expression in parietal cells (PCs) (white arrows, bottom right). Scale bars: 50 µm (top) and 20 µm (bottom) panels. b Corpus uptake of oleic acid is reduced in EC- Cd36 −/− mice. 16 h fasted 4-month-old mice were given a retro-orbital injection of [ 3 H]oleic acid and tissues collected for measuring radioactivity 5 min later. c Sum of cardiolipin species enriched with ω6 PUFA d Transmission EM showing EC- Cd36 −/− PCs lack the mitochondria (“M”)-free area near the plasma membrane bordering capillaries (right panel). Scale bar: 2 µm . e Quantification of mitochondrial circularity and aspect ratio (major/minor axis). f mRNA expression of gastric hormones, fibronectin, and inflammatory markers in corpus of EC- Cd36 −/− mice. g Fibronectin staining (green) in corpus of control and EC- Cd36 −/− mice, scale bar: 50 µm. Significance was calculated using a two-sided unpaired t test or a two-way ANOVA followed by post hoc tests by Sidak multiple comparisons ( c ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data are means + SEM. b n = 6; c n = 8–9; e n = 1500 mitochondria from 11 PCs/genotype; f n = 6.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Expressing, Injection, Radioactivity, Transmission Assay, Clinical Proteomics, Membrane, Staining, Control

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: Low CD36 expression associates with susceptibility to gastric disorders.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Expressing

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: a Genomic coordinates along chromosome 7q (x-axis), with negative logarithm of p-values (y-axis) for SNP association with gastrointestinal hemorrhage p = 7.9 -17 . Location of rs144921258 in CD36 on chr7q is indicated by a diamond. The schematic of the CD36 gene below the plot also depicts the position of GNAT3, which overlaps a distal promoter of CD36. b Position of rs144921258 relative to the abundant proximal CD36 promoter 1B, (shown with its long 3’ untranslated region) and to SNPs previously associated with methylation at CpG sites with impact on CD36 expression and lipid handling. Red CpGs: SNP-associated methylation sites that impact CD36 expression. Purple SNPs: SNPs associated with both lipid levels and low CD36 expression. See also Table and Results.

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

Techniques: Methylation, Expressing

Journal: Communications Biology

Article Title: CD36 maintains the gastric mucosa and associates with gastric disease

doi: 10.1038/s42003-021-02765-z

Figure Lengend Snippet: CD36 is abundant in corpus endothelial cells (EC) and expressed in parietal cells (PC). It is not detected in chief cells (CC), but its deficiency through vagal input and likely muscarinic receptor 1, suppresses CC release of leptin and gastric intrinsic factor (GIF), respectively. GIF is necessary for ileal vitamin B12 absorption. CD36 deletion reduces endothelial delivery of fatty acids to the corpus, tissue energy stores and mitochondrial FA oxidation, critical for stem cell functionality. Impaired renewal of the CD36 -/- gastric epithelium reflects defective PC progenitor differentiation to PCs and not progenitor proliferation or dysfunction of mature PCs. Low gastric leptin secretion could further impair tissue healing. The reduced tissue ability for adequate repair would contribute to the etiology of gastric disease, as supported by data from two biomedical databases (see Fig. and Table ).

Article Snippet: Tissue proteins separated by SDS-PAGE (4–12% acrylamide; Invitrogen), transferred to polyvinlyidene fluoride membranes (Millipore) and blocked (Li-COR Biosciences, Lincoln, NE), were incubated overnight (4 °C) with primary antibodies; mouse anti-mouse β-actin (1:5000), intrinsic factor (1:1000) (Santa Cruz Biotech), goat anti-mouse CD36 (1:1000, R&D Systems).

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