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: PLoS ONE

Article Title: Nanoparticles Containing Curcumin Useful for Suppressing Macrophages In Vivo in Mice

doi: 10.1371/journal.pone.0137207

Figure Lengend Snippet: Panel A, C, E, and G show results obtained from mice injected twice with control liposomes (200 μl) 48 and 72 h before analysis. Panel B, D, F, and H show those from mice injected with curcumin/liposome (200 μl, 1.8 mg/ml of curcumin concentration) in the same manner. In panel A and B, peritoneal cells were not stained; in panel C and D, were stained with Alexa Fluor 488 anti-CD90.2 (Thy1.2) antibody; in panel E and F, were stained with FITC anti-F4/80 antibody; in panel G and H, were stained with Alexa Fluor488 anti-CD36 antibody. Dots plotted in the Figures show fluorescence intensity. The vertical axis and the horizontal axis represent green red fluorescence intensity, respectively. GRN-HLog or RED-HLog means the highest fluorescence intensity on log scale. Excitation was performed with a blue laser at 488 nm. Green fluorescence was detected with a 525 ± 30 nm band pass filter and red fluorescence with a 680 ± 30 nm band pass filter. It is notable that dots in the circled areas of panel F and H apparently show higher green fluorescence compared to panel B and D.

Article Snippet: Peritoneal cells (1 × 10 7 /ml) were stained with Alexa Fluor 488 anti-mouse CD90.2 (Thy-1.2) Antibody(BioLegend)at 2.5 μg/ml, fluorescein isothiocyanate (FITC) rat anti-mouse F4/80(AbD serotec) at 10 μg/ml, Alexa Fluor 488 anti-mouse CD36 Antibody (BioLegend) at 2.5μg/ml, and PerCP/Cy5.5 anti-mouse/human CD11b (BioLegend) at 2.5 μg/ ml, respectively.

Techniques: Injection, Liposomes, Concentration Assay, Staining, Fluorescence

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 1. A, genomic structure of murine CD36, targeting vector, and mutated allele. The black boxes represent murine CD36 exons 1, 2, and 3 (left to right). The arrows below indicate transcription orientation. The bent arrow indicates the translation start site of CD36. Restriction endonuclease sites are designated as follows. X, XbaI; B, BamHI, H3, HindIII, N, NotI. The homologous sequences between the vector and the genomic locus are, for the 59 arm, just following the first HindIII site and just after exon 2; these sequences for the 39 arm are about 1 kb upstream of the third HindIII site until the 39-most BamHI site shown. The resulting mutated allele lacks exon 3, contains a neor cassette in the opposite transcriptional orientation, and is shorter by about 1 kb. No specific site of recombination should be inferred by this diagram. B, Southern blot analysis of genomic tail DNA. DNA was digested with HindIII, transferred to ZetaProbe GT, and hybridized with a 32P-la- beled probe, which is indicated in A. The restriction endonuclease and probe were chosen to differentiate between the wild type genomic locus, a random integration event, and a homologously recombined allele, on the basis of the size of the fragment to which the probe hybridized. Wild type (1/1), heterozygous (1/2), and null (2/2) animals are indicated and give the expected fragment sizes for the endogenous allele and a homologously recombined allele.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Plasmid Preparation, Southern Blot

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 2. A, immunoprecipitation/Western blot analysis of CD36. Pro- tein from fat was prepared by solubilization in radioimmune precipita- tion buffer containing protease inhibitors. Fat was chosen because of the high expression of CD36 in this tissue. CD36 was immunoprecipi- tated with a polyclonal antibody against the rat homologue of CD36 and then identified by SDS-PAGE followed by Western blotting with the same antibody. CD36 protein was detected in wild type (1/1) mice but was absent in null mice (2/2), as indicated by the 88-kDa band desig- nated with the arrowhead. B, flow cytometry analysis of buffy coat blood cells. Polyclonal anti-CD36 antibody, followed by a fluorescein-conju- gated second antibody, was used to detect CD36 protein expression on mononuclear cells from wild type mouse blood. Incubation with second antibody alone was similar to that of the null animal.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Immunoprecipitation, Western Blot, Expressing, SDS Page, Flow Cytometry, Incubation

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 3. Cell association of 125I-oxidized LDL to elicited perito- neal macrophages from wild type and CD36 null animals. Cells were incubated with increasing concentration of 125I-oxidized LDL for 5 h at 37 °C, washed, and solubilized as described. A significant amount of binding and internalization of oxidized LDL can be accounted for by CD36, as inferred from the decrease in cell association observed in cells from CD36 null animals. E and G, wild type; M and f, CD36 null. Open symbols demonstrate cell association in the presence of 50 mg/ml fucoidan.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Incubation, Concentration Assay, Binding Assay

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 4. Fatty acid transport into adipocytes of wild type and CD36 null animals. Data are from pooled adipocytes isolated from three animals per group per experiment and shown for fatty acid:BSA ratios of 1.5 (E, M) and 0.5 (G, f). Rates were obtained from five time measurements (0–120 s) and normalized to the DNA content as deter- mined on aliquots of the cell suspension. E and G, wild type; M and f, CD36 null. The mean 6 S.E. of triplicates is plotted. A representative experiment is shown. Inset, Hanes plot of fatty acid uptake by adipo- cytes as a function of unbound fatty acid concentration. The Hanes plot of S/v versus S, where S represents unbound fatty acid concentration in nM and v represents velocity or rate in nmol/min/mg of DNA, has its x intercept as the 2Km, and its slope is 1/Vmax. The data were derived from two experiments conducted at the different fatty acid:BSA ratios shown. In each experiment, complete time courses were performed for each ratio, and these were used to obtain the rates per minute. Un- bound fatty acid concentrations were calculated using a computer rou- tine as described previously (83) and were based on the fatty acid:BSA dissociation constants provided by Richieri and Klienfeld (89). Data were fitted by linear regression (wild type: r2 . 0.9). Data for CD36 null mice yields a line parallel to the x axis, so no Km could be obtained over the range of unbound concentrations used; the Km is apparently very high, or the process is not saturable. G, wild type; f, CD36 null.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Isolation, Suspension, Concentration Assay, Derivative Assay

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 5. Concentration of plasma glucose in wild type and CD36 null animals. Animals were fasted for 18 h, tail vein blood was col- lected into sodium fluoride, and glucose concentration was assayed. Bar, median value. *, p , 0.01.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Concentration Assay, Clinical Proteomics

Journal: The Journal of biological chemistry

Article Title: A null mutation in murine CD36 reveals an important role in fatty acid and lipoprotein metabolism.

doi: 10.1074/jbc.274.27.19055

Figure Lengend Snippet: FIG. 6. Fast protein liquid chromatography of lipoproteins from wild type and null animals. Plasma was collected by heart puncture from 4–6 age- and sex-matched animals of each genotype. Total cholesterol (TC; panel A), phosphatidylcholine (PC; panel B), and triacylglycerol concentration (TG; panel C) were measured in each fraction and are shown. An increase in cholesterol was apparent in all lipoprotein pools in the CD36 null animal, with the greatest increase present in the HDL fractions. HDL particles were notably larger and had an increase in phosphatidylcholine. There was a large increase in VLDL-associated triacylglycerol in CD36 null animals and an increase in intermediate density lipoprotein (IDL) particles.

Article Snippet: Materials—Cell culture reagents including leukemia-inhibitory factor (Esgro) were obtained from Life Technologies, Inc.; fetal bovine serum was from Gemini Bio-Sciences, Inc.; total cholesterol, triacylglycerol, albumin, and glucose quantitative assay kits and calibrators and miscellaneous chemicals were from Sigma; standards and a quantitative assay kit for serum nonesterified fatty acids were from Wako; FITC-conjugated goat anti-rabbit IgG was from Kirkegaard and Perry Laboratories; rabbit anti-rat CD36 IgG was prepared as described by Ryeom et al. (74); FITC-conjugated rat anti-mouse SR-A antibody (2F8) was from Serotec, Ltd.; carrier-free 125I-NaCl was from Amersham Pharmacia Biotech; [a-32P]dCTP, 14C-labeled palmitate, and 3H-labeled oleate were from NEN Life Science Products; the targeting vector, pPNT, was a gift from H.-C. Liou (Weill Medical College of Cornell University); the murine CD36 cDNA was a gift from G. Endemann (Scios Nova Inc.); CJ7 embryonic stem (ES) cells, passage 7, and neomycin-resistant (neor) mice were gifts of V. Soarez and E. Lacey (Sloan Kettering Institute); blastocyst injections were done through the core facilities at Sloan Kettering Institute and Rockefeller University; and fast protein liquid chromatography of plasma lipoproteins was performed in collaboration with the Rogosin Institute Clinical Research Laboratory for Comprehensive Lipid Analysis (Drs. T. Parker and D. Levine, Directors).

Techniques: Fast Protein Liquid Chromatography, Clinical Proteomics, Concentration Assay

Journal: ERJ Open Research

Article Title: The effects of exogenous lipid on THP-1 cells: an in vitro model of airway aspiration?

doi: 10.1183/23120541.00026-2016

Figure Lengend Snippet: PMA/THP-1 lipid uptake. PMA/THP-1 cells were incubated with 10% vol/vol Calogen for 24 h with and without inhibitors and were stained with ORO to detect lipid accumulation. a) Untreated cells were compared with b) cells treated with Calogen alone and c) in the presence of cytochalasin D and d) CD36 monoclonal blocking antibody. Inset images are magnified representative regions of each treatment. e) Cells treated with Calogen were also visualised under TEM. Scale bar=1 μm. f, g) Lipid droplets were identified and magnified to reveal dark stained remnants of lipid encased in membrane-bound vesicles, as indicated by the arrows. Scale bars=500 nm (f) and 100 nm (g).

Article Snippet: The plate was centrifuged (350 g , 3 min, 4 °C) and washed twice with PBS before incubating (30 min at 4 °C) with mouse monoclonal primary antibodies (CD11b, CD14, CD16, CD23, CD24, CD32, CD36, CD64, CD80, CD86, CD163, CD206 (20 μg·mL −1 , AbD Serotec, Oxford, UK) or CD32 [ ]).

Techniques: Incubation, Staining, Blocking Assay, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Aged Mouse Cortical Microglia Display an Activation Profile Suggesting Immunotolerogenic Functions

doi: 10.3390/ijms19030706

Figure Lengend Snippet: Expression of Cd206, Cd36 and Cd86 in F4/80 + cortical microglia. ( A ) Gating strategy and representative dot plots of Cd206 + , Cd36 + and Cd86 high cortical microglia from young (6 months) and aged (24 months) mice. Quantification of ( B ) F4/80 + , ( C ) Cd206 + , ( D ) Cd36 + and ( E ) Cd86 high microglia. Data are given as percentages of F4/80 + microglia ± SEM from three animals per age. p -values derived from student’s t -test are * p < 0.05, ** p < 0.01 and *** p < 0.001.

Article Snippet: Cells were stained with primary antibodies directed against Cd11b (1:20, 53-0112-82, eBiosciene, Thermo Fisher Scientific, Waltham, MA, USA), Cd206 (5 μL, FAB2535C, R&D Systems, Minneapolis, MN, USA), Cd36 (5 μL, MCA2748A647, AbD Serotech, BIO-Rad, Puchheim, Germany), Cd45 (1:20, 17-0451-82, eBiosciene), Cd86 (5 μL, MCA2463PE, AbD Serotech) and F4/80 (5 μL, MCA497A488, AbD Serotech) at 4 °C for 15 min. Fc (fragment crystallizable region) receptor blocking (TrueStain fcX, 101319, Biolegend, San Diego, CA, USA) was used to avoid unspecific antibody binding.

Techniques: Expressing, Derivative Assay

Journal: JCI insight

Article Title: Chikungunya virus infection disrupts lymph node lymphatic endothelial cell composition and function via MARCO.

doi: 10.1172/jci.insight.176537

Figure Lengend Snippet: Figure 2. WT CHIKV infection disrupts LEC marker expression and elicits infiltration of LN sinuses. (A–D) WT mice were mock inoculated (n = 3) or inoculated in the footpad with 1 × 103 PFU CHIKV 181/25 (n = 5) or WT CHIKV (n = 5), and the dLN was collected at 48 hours after infection. (A) LN sections stained for B220 (B cells, blue) and Lyve1 (LECs, white). Scale bar: 200 μm. (B) LN sections stained for B220 (B cells, blue) and MARCO (red). Scale bar: 200 μm. (C) Higher-magnification imag- es of subcapsular and medullary sinus regions in LNs stained for B220 (B cells, blue), Lyve1 (floor and medullary LECs, white), CD36 (ceiling LECs, green), and MARCO (red). Scale bar: 50 μm. (D) LNs stained for B220 (B cells, blue), ERTR-7 (fibroblasts, white), nuclei (red), and CD11b (myeloid cells, green). Scale bar: 200 μm (left), 50 μm (right). Images are representative of 3–5 dLNs per group (2 independent experiments).

Article Snippet: Sections were blocked with 5% goat, donkey, bovine, rat, or rabbit serum and then stained with 1 or more of the following antibodies: ERTR-7 (rat monoclonal, ERTR7, Abcam), B220 (RA3-6B2, Thermo Fisher Scientific), Lyve1 (ALY7, Thermo Fisher Scientific), CD11b (M1/70, BioLegend), CD36 (HM36), and/or MARCO (MCA1849, Serotec).

Techniques: Infection, Marker, Expressing, Staining

Journal: Journal of Neuroscience

Article Title: Microglial Dysfunction and Defective -Amyloid Clearance Pathways in Aging Alzheimer's Disease Mice

doi: 10.1523/jneurosci.0616-08.2008

Figure Lengend Snippet: Figure 2. Gene expression profile of adult WT CD11b microglia and CD11b neg cells (endothelial cells and astrocytes) mea- suredbyQPCR.A,ProtocolusedforisolatingCD11b cellsfromadultmousebrains.B,CD11b cells(blackbars)havehighlevels ofCD11bRNAandbarelydetectablelevelsofcellmarkersforastrocytes(GFAP)andendothelialcells(CD31)confirminghighpurity oftheCD11b cellspreparations.TheCD11b negpopulation(graybars)expressesCD31andGFAP,butnoCD11.C,CD11b cells isolated using our protocol expressed SRA, SRB1, and CD36 (3 known microglia receptors), some RAGE, and negligible levels of MARCO.

Article Snippet: APC-labeled anti-mouse CD11b (1 g/ml) (BD Biosciences Pharmingen) or APClabeled hamster anti-mouse CD36 clone HM36 (1 g/ml) (BioLegend) or Alexa647-labeled anti-CD204 (SRA) clone 2F8 (2.5 g/ml) (AbD Serotec) or isotype-matched control antibodies (same concentrations as primary antibodies) were added and incubated on ice for 1 h. Cells were then fixed by the addition of 2% PFA.

Techniques: Gene Expression, Isolation

Journal: Journal of Neuroscience

Article Title: Microglial Dysfunction and Defective -Amyloid Clearance Pathways in Aging Alzheimer's Disease Mice

doi: 10.1523/jneurosci.0616-08.2008

Figure Lengend Snippet: Figure 3. Reduced expression of A-binding receptors in microglia from old transgenic PS1-APP mice. Expression of A- bindingreceptorsinCD11b cellswascomparedbetweentransgenicPS1-APPmiceandtheirage-matchedWTlittermatesat1.5, 3,8,and14monthsofage.DatarepresenttheaveragevaluesobtainedbyQPCRonsixsetsofmicefor3and14monthsofageand onfivesetsofmiceat1.5and8monthsofage.*Valuesofpareshownonlyifdifferencesaresignificant(0.05).A,B,D,At8and 14monthsofage,PS1-APPtransgenicmiceshowsignificantlyreducedexpressionofSRA(A),CD36(B),andRAGE(D)compared withtheirWTlittermates.C,TherearenosignificantdifferencesinexpressionofSRB1observedbetweentransgenicandWTmice at any age. E, There was negligible expression of MARCO. Error bars indicate SEM.

Article Snippet: APC-labeled anti-mouse CD11b (1 g/ml) (BD Biosciences Pharmingen) or APClabeled hamster anti-mouse CD36 clone HM36 (1 g/ml) (BioLegend) or Alexa647-labeled anti-CD204 (SRA) clone 2F8 (2.5 g/ml) (AbD Serotec) or isotype-matched control antibodies (same concentrations as primary antibodies) were added and incubated on ice for 1 h. Cells were then fixed by the addition of 2% PFA.

Techniques: Expressing, Binding Assay, Transgenic Assay

Journal: Journal of Neuroscience

Article Title: Microglial Dysfunction and Defective -Amyloid Clearance Pathways in Aging Alzheimer's Disease Mice

doi: 10.1523/jneurosci.0616-08.2008

Figure Lengend Snippet: Figure 6. Treatment of N9 cells with TNF decreases expression of SRA and CD36 and re- duces uptake of A. A, B, N9 microglia-like cells were treated with TNF overnight, and ex- pression of SRA and CD36 were measured by QPCR (A) and by cell surface staining and flow cytometry (B). Data represent the average of five separate experiments and are displayed for treatedcellsaspercentageexpression(%)ofuntreatedcontrols(100%).B,Farright,Resultsof TNF- treatment on binding/uptake of A-labeled with Hylite fluor-488. TNF--treated or untreated N9 cells were incubated with fluorescent A for 2 h, and then cell-associated fluo- rescence was measured by flow cytometry. Data represent an average of five experiments and are displayed for TNF-treated cells as percentage (%) of mean fluorescence intensity of un- treated cells. Error bars indicate SEM.

Article Snippet: APC-labeled anti-mouse CD11b (1 g/ml) (BD Biosciences Pharmingen) or APClabeled hamster anti-mouse CD36 clone HM36 (1 g/ml) (BioLegend) or Alexa647-labeled anti-CD204 (SRA) clone 2F8 (2.5 g/ml) (AbD Serotec) or isotype-matched control antibodies (same concentrations as primary antibodies) were added and incubated on ice for 1 h. Cells were then fixed by the addition of 2% PFA.

Techniques: Expressing, Staining, Flow Cytometry, Binding Assay, Labeling, Incubation, Fluorescence