Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Anti-atherogenic effects of CD36-targeted epigallocatechin gallate-loaded nanoparticles

doi: 10.1016/j.jconrel.2019.04.018

Figure Lengend Snippet: The binding and uptake of NBD-labeled Vnano and L-Vnano by mouse peritoneal macrophages via their CD36 receptor. A, Mouse peritoneal macrophages were treated with non-targeted Vnano or targeted L-Vnano at 37°C; B, Mouse peritoneal macrophages were treated with Vnano or L-Vnano at 4°C; C, Mouse peritoneal macrophages were treated with red RPE-labeled anti-mouse CD36 antibody (λ of excitation is 496 nm, λ of emission is 578 nm) in combination with Vnano or L-Vnano at 4°C; D, Mouse peritoneal macrophages were transfected without CD36 siRNA (control) or with scramble siRNA (negative control) or with CD36 siRNA (CD36 knockdown). CD36 siRNA transfection decreased CD36 protein expression in mouse peritoneal macrophages; E, The binding and uptake of L-Vnano by control, CD36 negative control or CD36 knockdown mouse peritoneal macrophages. Bar length was 200 μm. NBD-labeled Vnano and L-Vnano were green (λ of excitation is 460 nm; λ of emission is 535 nm). Cell nuclei were stained by DAPI (λ of excitation is 358 nm, λ of emission is 461 nm) (blue color). Images were representatives of three independent experiments.

Article Snippet: Moreover, for competitive binding assay with mouse CD36 antibody, mouse peritoneal macrophages were treated with RPE-labeled anti-CD36 antibody (MCA2748PE, AbD Serotec, Raleigh, NC, USA) in combination with NBD-labeled Vnano or L-Vnano at 4°C for 2 hours.

Techniques: Binding Assay, Labeling, Transfection, Negative Control, Expressing, Staining

Journal: Cell reports

Article Title: Neutral ceramidase-dependent regulation of macrophage metabolism directs intestinal immune homeostasis and controls enteric infection

doi: 10.1016/j.celrep.2022.110560

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CD36 (147 Sm) , Fluidigm , Cat# 3147018B;RRID:N/A.

Techniques: Recombinant, Western Blot, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Cell Culture, IP Phosphatase Assay, Software

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

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Reduced hematoma absorption and aggravated neurologic deficits in CD36-deficient patients with ICH. (A) Typical cranial CT images of CD36-normal (n = 11) and CD36-deficient (n = 11) ICH patients. Original magnification ×10. (B) Comparison of hematoma volumes in the CD36-deficient and CD36-normal groups at admission and 7 d after the onset of ICH. *p < 0.05 versus the CD36-normal group. (C) Decreased absorption rate in the CD36-deficient patients relative to the normal group. **p < 0.01 versus the CD36-normal group. (D) NIHSS scores and (E) mRS scores for ICH patients. *p < 0.05 versus the CD36-normal group, **p < 0.01 versus the CD36-normal group.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques:

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Enhanced CD36 expression in perihematomas following ICH. (A) Detection of CD36 mRNA expression in mice subjected to ICH using real-time quantitative RT-PCR. The data are expressed as fold increases relative to naive animals. **p < 0.01 versus sham, n = 6. (B) Detection of CD36 protein expression in mice subjected to ICH using Western blot. **p < 0.01 versus sham, n = 6. (C) Detection of CD36 expression in astrocytes, neurons, and microglia in the perihematomal tissues of mice using flow cytometry at 3 d after ICH. The left panel is the representative flow cytometry plot and the lower right panel is the percentage of CD36+ cells in neurons, astrocytes, and microglia. Microglia were marked by CD45intCD11b+, whereas neurons were identified as β-III tubulin+, and astrocytes were identified as GFAP+. **p < 0.01 versus sham, ##p < 0.01 versus astrocytes or neurons, n = 3. (D) Detection of CD36 expression in human perihematomal tissues of ICH patients using fluorescence immunohistochemistry; CD36 expression was labeled with a CD36 Ab (red), astrocytes, neurons, and microglia were labeled with GFAP (green), Neun (green), and Iba-1 (green), respectively. The merged images of the overlay of CD36 together with astrocytes, neurons, and microglia were shown as yellow, and the nuclei were stained with DAPI (blue). The arrows indicate positive cells. Scale bars, 80 μm.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Flow Cytometry, Fluorescence, Immunohistochemistry, Labeling, Staining

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: CD36-mediated hematoma absorption in mice with ICH. (A) Serial coronal sections of mouse brain tissues 5 d after the onset of ICH. Original magnification ×1.5. (B) Comparison of hematoma volumes between WT and CD36−/− mice. The data in the left and right panels are the Image-Pro Plus and hemoglobin detection results, respectively. **p < 0.01 versus the WT group, n = 6. (C) The NDS of WT and CD36−/− mice at 1, 3, 5, and 7 d after the onset of ICH. *p < 0.05 versus the WT group at the corresponding time points, n = 6. (D) Brain water content and (E) mRNA expression of inflammatory factors 3 d after the onset of ICH. The data in (E) were assessed by real-time quantitative RT-PCR and were expressed as fold increases relative to naive animals. *p < 0.05 versus the WT group; **p < 0.01 versus the WT group, n = 6. Cereb, cerebellum; Cont BG, contralateral basal ganglia; Cont CX, contralateral cortex; Ipsi BG, ipsilateral basal ganglia; Ipsi CX, ipsilateral cortex.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Quantitative RT-PCR

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: CD36-mediated engulfment of RBCs by microglia. (A) Observation of the phagocytosis of RBCs (CFSE-labeled, green) by microglia (Alexa Fluor 594–labeled, red) using confocal microscopy. The nuclei were stained with Hoechst. Serial sections along the z-axis were acquired and compiled as images. The x–z-axis is below, and the y–z-axis is on the right. The arrows indicate the RBCs engulfed by microglia. Scale bars, 40 μm. Detection of changes in the phagocytic ability of microglia using flow cytometry is shown. Microglia were stained with anti-mouse CD11b-PE and RBCs were stained with CFSE. (B) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel). (C) Average fluorescence intensity of CFSE in the PE+CFSE+ microglia. (D) The number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the WT microglia group, n = 6 for all graphs.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Labeling, Confocal Microscopy, Staining, Flow Cytometry, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: TLR4 signaling downregulates CD36 expression and increases hematoma absorption. (A) Detection of CD36 expression in perihematomal tissues 3 d after the onset of ICH using Western blot. **p < 0.01 versus the WT group, n = 6. (B) Detection of microglia CD36 expression using flow cytometry. **p < 0.01 versus the WT group, n = 6. (C) Five days after the onset of ICH, the hematoma volume was smaller in MyD88−/− and TLR4−/− mice than in WT mice. Measurement of hematoma volume using Image-Pro Plus (left panel) and hemoglobin detection (right panel) are shown. **p < 0.01 versus the WT group, n = 6. Detection of changes in the phagocytic capacity of microglia using flow cytometry is depicted. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (D) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel); (E) average fluorescence intensity of CFSE in PE+CFSE+ microglia; (F) number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the WT microglia group, ##p < 0.01 versus the groups with anti-CD36 Abs, n = 6 for all flow cytometry graphs.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: Effects of TNF-α, IL-1β, and IL-10 on CD36 expression and phagocytosis in microglia. (A) Detection of changes in microglial CD36 protein levels using Western blot. *p < 0.05 versus the vehicle group, **p < 0.01 versus the vehicle group, n = 6. (B) Detection of microglial CD36 expression using flow cytometry. **p < 0.01 versus the vehicle group, n = 6. Detection of changes in the phagocytic ability of microglia using flow cytometry. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (C) Representative flow cytometry plot (left panel) and the percentage of PE+CFSE+ microglia that had engulfed RBCs (right panel). **p < 0.01 versus the vehicle group, n = 6. (D) Average fluorescence intensity of CFSE in PE+CFSE+ microglia. **p < 0.01 versus the vehicle group, n = 6. (E) Number of nonphagocytosed RBCs in the supernatant. *p < 0.05 versus the vehicle group, **p < 0.01 versus the vehicle group, n = 6.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence

Journal: The Journal of Immunology Author Choice

Article Title: CD36-Mediated Hematoma Absorption following Intracerebral Hemorrhage: Negative Regulation by TLR4 Signaling

doi: 10.4049/jimmunol.1400054

Figure Lengend Snippet: TAK-242 (TAK) increases CD36 expression and promotes hematoma absorption. (A) Detection of CD36 protein expression using Western blot. **p < 0.01 versus the vehicle group, #p < 0.05 versus the TAK group, ##p < 0.01 versus the TAK group, n = 6. (B) Detection of microglia CD36 expression using flow cytometry. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. Detection of changes in the phagocytic capacity of microglia using flow cytometry. Microglia were stained with anti-mouse CD11b-PE, and RBCs were stained with CFSE. (C) A representative flow cytometry plot (left panel) displaying the percentage of PE+CFSE+ microglia (right panel) that had engulfed RBCs. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (D) The average fluorescence intensity of CFSE in PE+CFSE+ microglia. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (E) The number of nonphagocytosed RBCs in the supernatant. **p < 0.01 versus the vehicle group, ##p < 0.01 versus the TAK group, n = 6. (F) Detection of microglial catalase expression using real-time quantitative RT-PCR. The data are expressed as fold increases relative to the vehicle group. **p < 0.01, n = 6. (G) The changes in H2O2 content in the supernatant were measured. The results are presented as fold changes relative to vehicle group. **p < 0.01, n = 6. (H) Changes in hematoma volume in mice at 5 d after the onset of ICH. Measurement of hematoma volume using Image-Pro Plus (left panel) and detection of hemoglobin (right panel). **p < 0.01 versus the ICH plus vehicle group, #p < 0.05 versus the ICH plus TAK plus TNF-α group, n = 6.

Article Snippet: The membranes were incubated with mouse anti-human CD36 (for human blood, 1:400; Wako Chemicals USA, Richmond, VA) or rabbit anti-mouse CD36 (for perihematomal tissues and cells, 1:400; Abcam) at 4°C overnight; GAPDH (1:400; Santa Cruz Biotechnology, Dallas, TX) was used as a loading control.

Techniques: Expressing, Western Blot, Flow Cytometry, Staining, Fluorescence, Quantitative RT-PCR