Journal: JCI Insight

Article Title: Whole-body deletion of Endospanin 1 protects from obesity-associated deleterious metabolic alterations

doi: 10.1172/jci.insight.168418

Figure Lengend Snippet: ( A ) Detection of Flag-CD36 in 6Myc-Endo1 immunoprecipitate from HEK293T cells coexpressing 6Myc-Endo1 and Flag-CD36 compared with HEK293T cells transfected with Flag-CD36 alone. Representative blots of 3 independent experiments. ( B ) Detection of 6Myc-Endo1 after Flag-CD36 immunoprecipitation from HEK293T cells coexpressing 6Myc-Endo1 and Flag-CD36 compared with HEK293T cells transfected with 6Myc-Endo1 alone. Representative blots of 3 independent experiments. ( C ) Kinetics of Endo1 and CD36 protein expression in differentiated adipocytes during differentiation of adipocyte precursors isolated from the stromal vascular fraction of the s.c. adipose tissue of WT mice into mature white adipocytes. Results are expressed as mean ± SEM of 8 independent experiments. One representative Western blot is shown. ( D ) Confocal immunofluorescence detection and colocalization of Endo1 (rabbit anti-Endo1) and CD36 (goat anti-CD36) in differentiated adipocytes. Nuclei (blue) are stained with fluorescent DAPI dye. Scale bar: 20 μm. Representative images of 4 independent experiments. ( E ) Detection of endogenous Endo1 and CD36 after immunoprecipitation with Endo1 or CD36 antibodies from lysates of visceral adipose tissue (VAT), gonadal adipose tissue (GAT), and s.c. adipose tissue (SAT) of WT and KO mice. Endo1-KO adipocytes and tissues were used as negative controls. The molecular weights of protein markers are indicated (kDa). Representative blots of 2 independent experiments.

Article Snippet: Next, cells were incubated with primary Endo1 (1:500) (generated as described previously; ref. ) and primary CD36 (1:1,000) (RD Systems, catalog AF2519), primary Endo1 (1:500) and primary 53K (1:1,000, Abcam, catalog ab27043), or primary GM130 (1:1,000, Abcam, catalog ab169276) antibodies overnight at 4°C.

Techniques: Transfection, Immunoprecipitation, Expressing, Isolation, Western Blot, Immunofluorescence, Staining

Journal: JCI Insight

Article Title: Whole-body deletion of Endospanin 1 protects from obesity-associated deleterious metabolic alterations

doi: 10.1172/jci.insight.168418

Figure Lengend Snippet: ( A ) Cell surface expression of CD36 in mature white adipocytes. ** P < 0.01 versus WT. Results are expressed as mean ± SEM ( n = 4). Two-tailed t test. ( B ) Total CD36 expression in gonadal adipose tissue (GAT) and s.c. adipose tissue (SAT) . The molecular weights of protein markers are indicated (kDa). Results are expressed as mean ± SEM ( n = 5). ( C ) Immunofluorescence images of CD36 cell surface expression in differentiated white adipocytes (left panel). Level of cellular fluorescence determined by corrected total cell fluorescence per area (CTCF/area). Results are expressed as mean ± SEM ( n = 9). *** P < 0.005 versus WT. Two-tailed t test (right panel). Scale bar: 20 μm. ( D ) Lipid uptake in differentiated adipocytes, mature adipocytes, and differentiated myotubes. Results are expressed as mean ± SEM ( n = 5–6). * P < 0.05; ** P < 0.01; **** P < 0.001 versus basal. † P < 0.05; †† P < 0.01; †††† P < 0.001 versus WT. One-way ANOVA with Bonferroni correction.

Article Snippet: Next, cells were incubated with primary Endo1 (1:500) (generated as described previously; ref. ) and primary CD36 (1:1,000) (RD Systems, catalog AF2519), primary Endo1 (1:500) and primary 53K (1:1,000, Abcam, catalog ab27043), or primary GM130 (1:1,000, Abcam, catalog ab169276) antibodies overnight at 4°C.

Techniques: Expressing, Two Tailed Test, Immunofluorescence, Fluorescence

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Replacing saturated fat with unsaturated fat in western diet reduces foamy monocytes and atherosclerosis in male Ldlr –/– mice

doi: 10.1161/ATVBAHA.119.313078

Figure Lengend Snippet: EVOND compared to WD reduced foamy monocyte formation and inflammation in Ldlr–/– mice. A, Monocyte frequency in total leukocytes of mice on ND, WD, and EVOND (n=12–20/group). B, Side scatter (SSC) value and Nile red mean fluorescence intensity (MFI) of circulating monocytes of mice on different diets (3 months on diets for Nile red staining). C, Representative fluorescence-activated cell sorter (FACS) examples showing foamy monocytes in blood of Ldlr–/– mice on different diets (left panel). Monocytes (CD115+) were divided into two subsets based on CD36. Elevations in SSC indicated lipid accumulation and foamy monocyte formation; quantification of SSC values of CD36– and CD36+ monocytes in Ldlr–/– mice on diets (n=9–18/group; right panel). D, CD11c expression on CD36– and CD36+ monocytes in Ldlr–/– mice on diets (n=9–18/group). E, Expression of TNFα and IL-1β in monocytes of Ldlr–/– mice on diets (n=4/group). Data are shown as mean±SEM. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: 21 Antibodies and fluorescence-activated cell sorter (FACS) analysis of circulating monocytes For FACS analysis, monoclonal antibodies against the following mouse antigens were used: CD115 (PE, AFS98, eBioscience), CD204 (FITC, 2F8, Bio-Rad Laboratories), CD11c (PerCP-Cy5.5, N418, eBioscience), CD36 (FITC, MF3, Bio-Rad Laboratories), Ly-6C (APC, HK1.4, eBioscience), TNFα (PE, MP6-XT22, eBioscience), and IL-1β (PE, NJTEN3, eBioscience).

Techniques: Fluorescence, Staining, Expressing

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Replacing saturated fat with unsaturated fat in western diet reduces foamy monocytes and atherosclerosis in male Ldlr –/– mice

doi: 10.1161/ATVBAHA.119.313078

Figure Lengend Snippet: EVOND reduced monocyte CD36 expression and oxidized LDL (oxLDL) uptake. A, SSC value of monocytes (from ND-fed Ldlr–/– mice) after incubation with triglyceride-rich lipoprotein (TGRL) fraction from mice on WD or EVOND (n=6/group). B, Expression level of CD36 on circulating monocytes from mice on different diets (left panel; n=9–15 mice/group) or on monocytes from ND-fed Ldlr–/– mice after incubation with TGRL fraction from mice on WD or EVOND (right panel; n=6/group). C, Monocyte uptake of DiI-oxLDL. Total leukocytes from Ldlr–/– mice on WD or EVOND were incubated ex vivo with DiI-oxLDL in the absence or presence of anti-mouse CD36 antibody for 1 h, and DiI signals representing monocyte uptake of oxLDL were examined by FACS (n=4/group). Data are shown as mean±SEM. *p<0.05, ***p<0.001.

Article Snippet: 21 Antibodies and fluorescence-activated cell sorter (FACS) analysis of circulating monocytes For FACS analysis, monoclonal antibodies against the following mouse antigens were used: CD115 (PE, AFS98, eBioscience), CD204 (FITC, 2F8, Bio-Rad Laboratories), CD11c (PerCP-Cy5.5, N418, eBioscience), CD36 (FITC, MF3, Bio-Rad Laboratories), Ly-6C (APC, HK1.4, eBioscience), TNFα (PE, MP6-XT22, eBioscience), and IL-1β (PE, NJTEN3, eBioscience).

Techniques: Expressing, Incubation, Ex Vivo

Journal: Frontiers in Immunology

Article Title: Meningeal Foam Cells and Ependymal Cells in Axolotl Spinal Cord Regeneration

doi: 10.3389/fimmu.2019.02558

Figure Lengend Snippet: CD36 and TLR4 in regenerating cord in vivo and in vitro . (A) The scavenger receptor CD36 was detected on the endfeet of ependymal cells in control cord paraffin cross-sections. Fluorescence image. (B) CD 36 is also present on ependymal cell bodies in intact adult Axolotl spinal cord. Fluorescence image. (C) In paraffin sections from regenerating cord stump CD36 was detected in ependymal endfeet in the reactive meninges. Fluorescence image. (D) CD36 also present on ependymal cell bodies in regenerating cord, proximal stump. Fluorescence image. (E) In vitro , CD36 is found on foamy macrophages from 10D regenerate spinal cords 6 days in vitro (yellow arrows). Orange arrows show three nuclei in CD36 + MNGC. Inset in (E) shows a CD36 + MNGC with six nuclei. (F) CD36 + Ependymal cells (white arrows) in explant cultures from a 10D regenerate spinal cord 6 days in vitro . (G) TLR4 was detected in foamy macrophages (yellow arrows) in culture on cells from 14 days regenenerates, 17 days in vitro . (H) Ependymal cells from 14 days regenenerates, 17 days in vitro are also TLR4 + (white arrows). D, day; Regen, regenerating; MNGC, multinucleated giant cells; TLR4, toll like receptor 4. Magnification bar is shown in the lower portion of each image.

Article Snippet: CD36 antibody (R&D Systems MAB25191) was diluted to 2.5 ug/ml.

Techniques: In Vivo, In Vitro, Control, Fluorescence

Journal: Frontiers in Immunology

Article Title: Meningeal Foam Cells and Ependymal Cells in Axolotl Spinal Cord Regeneration

doi: 10.3389/fimmu.2019.02558

Figure Lengend Snippet: CD36 inhibition reduces DiI-Ox-LDL uptake by foamy macrophages. Seventeen days regenerate spinal cord regenerate explants, were treated with DiI-Ox-LDL at 10DIV for 24 h as the control (A) or with the CD36 inhibitor sulfo-N-succinimidyl oleate added at 10DIV plus DiI-Ox-LDL added at 11DIV for 24 h. (B) Inhibitor-treated explants showed less uptake of DiI-Ox-LDL in comparison to control. D, day; DIV, days in vitro ; Ox-LDL, oxidized low-density receptor; inhib, inhibitor; Regen, regeneration. Magnification bar is shown in the lower portion of each image.

Article Snippet: CD36 antibody (R&D Systems MAB25191) was diluted to 2.5 ug/ml.

Techniques: Inhibition, Control, Comparison, In Vitro

Journal: Respiratory Research

Article Title: An investigation of the resolution of inflammation (catabasis) in COPD

doi: 10.1186/1465-9921-13-101

Figure Lengend Snippet: Representative examples of different macrophage immunostaining intensities (absent/moderate vs. intense staining) for CD44, CD36, VEGF and TGFβ. For further explanations, see text.

Article Snippet: Formalin-fixed paraffin-embebbed tissue sections (3 μm thick) were immunostained with the following monoclonal mouse antibodies: anti-human CD44, Phagocytic Glycoprotein-1, clone DF1485 (Dako, Glostrup, Denmark); anti-human VEGF, clone VG1 (Dako, Glostrup, Denmark); anti-human TGFbeta (AbDSerotec, Oxford, UK); anti-human CD36, clone SMO, (AbDSerotec, Oxford, UK).

Techniques: Immunostaining, Staining

Journal: Respiratory Research

Article Title: An investigation of the resolution of inflammation (catabasis) in COPD

doi: 10.1186/1465-9921-13-101

Figure Lengend Snippet: Individual and mean (bars) values of the proportion of macrophages with intense staining for CD44 , CD36, VEGF and TGFβ in patients with COPD, smokers with normal spirometry and non-smokers. (S = current smokers; EX-S = former smokers; NS = non-smokers). For further explanations, see text.

Article Snippet: Formalin-fixed paraffin-embebbed tissue sections (3 μm thick) were immunostained with the following monoclonal mouse antibodies: anti-human CD44, Phagocytic Glycoprotein-1, clone DF1485 (Dako, Glostrup, Denmark); anti-human VEGF, clone VG1 (Dako, Glostrup, Denmark); anti-human TGFbeta (AbDSerotec, Oxford, UK); anti-human CD36, clone SMO, (AbDSerotec, Oxford, UK).

Techniques: Staining