Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 4. No protective effect of SS31 on GSH level and infarct size in CD36 KO mice. CD36 KO mice were subjected to 30 min MCAO. A, mice were treated with either saline (Veh) or SS31 (2 mg/kg body weight) immediately after reper- fusion, and GSH measurement was performed at 6-h post-ischemia. Values for GSHmeasurementareexpressedaspercentGSHdepletionintheipsilateralside compared with contralateral side. Error bars indicate S.D. (n 4 per group). B, mice were treated with either vehicle or SS31 (2 mg/kg body weight) immedi- ately after reperfusion, 6, 24, and 48 h. Infarct volumes were measured at 72-h post-ischemia. Error bars indicate S.D. (n 7 per group). No difference was observed between vehicle- and SS31-treated groups.

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Saline

Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 5. Effect of SS31 on ischemia-induced CD36 expression. C57BL/6 mice were subjected to 30 min MCAO and treated with saline (Veh) or SS31 (5 mg/kg body weight) immediately after reperfusion and again 6 h after ische- mia. A, for CD36 gene expression, total RNA was prepared from both hemi- spheres 24 h after ischemia, and CD36 mRNA level was determined. Error bars indicate S.D. (n 7). *, p 0.05 versus contralateral side; #, p 0.05 versus ipsilateral vehicle-treated group, one-way ANOVA with post-hoc Newman- Kuels test. B, correlation analysis of CD36 protein levels and infarct size. CD36 protein levels from SS31-treated MPM were expressed as arbitrary units. SS31 (5 mg/kg) was given at 0, 6, 24, and 48 h, and MPM were harvested 72 h after ischemia. Infarct volume was determined at 72 h after ischemia. CD36 protein levels were normalized against -actin levels. Note that CD36 protein level is positively correlated with infarct size (r 0.6390, p 0.0055).

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Expressing, Saline, Gene Expression

Journal: Journal of Biological Chemistry

Article Title: A Novel Cell-permeable Antioxidant Peptide, SS31, Attenuates Ischemic Brain Injury by Down-regulating CD36

doi: 10.1074/jbc.m609388200

Figure Lengend Snippet: FIGURE 6. Effect of SS31 on oxLDL-induced CD36 expression in MPM. Thioglycolate-elicited MPM were cultured and incubated with saline, oxLDL, andSS31peptidessinglyorincombinationasindicated.A,CD36mRNAlevels were determined 48 h after treatments. Error bars indicate S.D. (n 6 per group) *, p 0.05 versus V; #, p 0.05 versus oxL. B, CD36 protein levels were determined 48 h after treatments by Western blot analysis. Experiments were performed four times (n 4 per group). For each set of experiment, CD36 protein band densities were normalized against -actin. A vehicle-treated blot was used as a reference standard (100%), and CD36 band intensity was calculated based on the density of vehicle-treated blot. H, heart (positive control); KO, CD36 KO brain (negative control); Veh, saline; oxL, 25 g/ml oxLDL; SH, 106 M SS31; SL, 108 M SS31; #, p 0.05 versus oxL, one-way ANOVA with post-hoc Newman-Kuels test.

Article Snippet: Filters were treated for 1 h in TBS (pH 7.2) containing 0.1% Tween-20 and 5% dry milk, and then incubated with CD36 polyclonal antibodies (1:1000; AF2519; R&D Systems, Minneapolis, MN) followed by mouse IgA-HRP (Sigma).

Techniques: Expressing, Cell Culture, Incubation, Saline, Western Blot, Positive Control, Negative Control

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