Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + orients naïve macrophages and reprograms TAMs towards an M1-like profile. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). ( A ) The proportion of macrophages expressing M1/M2 membrane markers (CD32, CD64, CD80, TLR2, CD163, CD36 and CD206) was determined by flow cytometry after three days of co-culture with SKOV3-R2 + cells. ( B ) The release of cytokines (IL1β, IL12, TNFα, IL6, IFNγ, IL10) and chemokines (CCL2, CCL4, CCL5, CXCL9 and CXCL10) in the culture medium was determined by AlphaLISA after three days of co-culture with SKOV3-R2 + cells. Data shown (boxplots) are the results from three different experiments (performed with three different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD36-PEVio770 , Miltenyi , 130-110-742 , REA770 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Expressing, Membrane, Flow Cytometry, Co-Culture Assay

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: Scientific Reports

Article Title: Endothelial Cell Phenotypes are Maintained During Angiogenesis in Cultured Microvascular Networks

doi: 10.1038/s41598-018-24081-z

Figure Lengend Snippet: Illustration of CD36 labeling in ex vivo cultured microvascular networks and stimulated in vivo microvasculature. Comparison between Day 3 ( Ex Vivo ) ( A–C ) and Day 3 ( In Vivo ) ( D–F ) microvascular networks revealed diminishing CD36 labeling along the length of the capillary sprout becoming absent at the tip. Arrows identify examples of CD36-negative capillary sprouts. Scale bars = 200 µm. ( G , H ) Capillary sprouts exhibited a substantial decrease in CD36 labeling compared to larger network vessels ( > 10 µm) in Day 3 ( Ex Vivo ) and Day 3 ( In Vivo ) tissues. White and black bars represent “Sprouts” and “Networks” respectively for Day 3 ( Ex Vivo ) and Day 3 ( In Vivo ) groups. The *** indicates a significant difference of p < 0.001 by Mann-Whitney U test.

Article Snippet: The following primary antibodies were used: UNC5b (1:100; Abcam; Cambridge, MA), VEGFR-2 (1:50; Santa Cruz Biotechnology; Dallas, TX), Alexa-568 Phalloidin (1:50; Invitrogen; Carlsbad, CA), CD36 (1:100; Novus Biologicals; Littleton, CO), and PECAM (1:200; BD Pharmingen; San Jose, CA).

Techniques: Labeling, Ex Vivo, Cell Culture, In Vivo, Comparison, MANN-WHITNEY

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