rat antimouse cd36 alexa fluor647  (Bio-Rad)

Journal: Journal of Lipid Research

Article Title: Foamy monocytes and atherogenesis in mice with combined hyperlipidemia and effects of antisense knockdown of apoCIII

doi: 10.1016/j.jlr.2025.100763

Figure Lengend Snippet: Changes in monocyte and subset phenotypes in Ldlr −/− apoCIIItg mice. Ldlr −/− ApoCIIItg and Ldlr −/− mice were fed WD for 6 weeks or as indicated otherwise. A: Change in CD11c MFI of foamy (CD11c + ) monocytes (n = 8–10 mice/group), CD11c MFI of foamy (CD11c + ) monocytes, and monocyte adhesion to VCAM-1 examined by ex vivo flow adhesion assay. B: Monocyte CD36 MFI and OxLDL uptake ex vivo. Data show representative FACS examples and quantification of OxLDL uptake by CD11c + monocytes. C: Representative FACS examples and quantification of intracellular TNFα and IL-1β in CD11c + and CD11c – monocytes in Ldlr −/− ApoCIIItg mice. D: Intracellular TNFα and IL-1β in CD11c + monocytes in Ldlr −/− ApoCIIItg and Ldlr −/− mice. E: CX3CR1 MFI on CD11c + and CD11c – monocytes in mice. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 compared to Ldlr −/− group (A).

Article Snippet: Monocyte phenotypes were analyzed by FACS using monoclonal antibodies (mAbs) to mouse antigens, including CD115 (PE, AFS98, ThermoFisher Scientific, Waltham, MA; or BV421, AFS98, BioLegend, San Diego, CA), CD11c (PerCP-Cy5.5, N418, ThermoFisher Scientific; or APC or FITC, N418, BioLegend), CD36 (FITC, MF3, Bio-Rad Laboratories, Hercules, CA), CD204 (FITC, 2F8, Bio-Rad Laboratories), Ly-6C (APC, HK1.4, ThermoFisher Scientific), CX3CR1 (FITC, SA011F11, BioLegend), MHCII (I-A/I-E; PE, M5/114, BD Biosciences, San Jose, CA), CD43 (PE, S11, BioLegend), TREML4 (PE, 16E5, BioLegend), XCR1 (PE, ZET, BioLegend), tumor necrosis factor α (TNFα; PE, MP6-XT22, ThermoFisher Scientific), and interleukin-1β (IL-1β; PE, NJTEN3, ThermoFisher Scientific), with appropriate isotype-negative controls.

Techniques: Ex Vivo, Cell Adhesion Assay

Journal: Journal of Lipid Research

Article Title: Foamy monocytes and atherogenesis in mice with combined hyperlipidemia and effects of antisense knockdown of apoCIII

doi: 10.1016/j.jlr.2025.100763

Figure Lengend Snippet: Monocyte uptake of TGRL and phenotypic changes. A: Representative FACS examples of mouse monocyte uptake of TGRL in vivo and ex vivo. Left panel (in vivo): DiI-mTGRL isolated from Ldlr −/− ApoCIIItg mice and labeled with DiI were intravenously injected into WT mice (recipients), and monocyte uptake of DiI-mTGRL was examined in recipient blood 24 h later after staining for CD204 and CD11c. Right panel (ex vivo): Blood from WT mice was incubated with or without DiI-mTGRL for 3 h, and monocyte uptake of DiI-mTGRL was examined after staining for CD204 and CD11c. Data shown were gated monocytes (CD204 + ). B: Representative FACS examples of THP-1 monocyte uptake of TGRL (indicated by DiI signal after incubation with DiI-TGRL for 4 h) and lipid accumulation (indicated by elevated SSC and Nile Red staining after incubation with TGRL for 48 h). C: Monocyte uptake of TGRL and lipid accumulation with lipoprotein lipase (LPL) inhibition or addition. THP-1 monocytes or blood from healthy humans were incubated with DiI-labeled or unlabeled postprandial TGRL in the presence or the absence of orlistat (orli) for 4 h (upper panels for DiI-TGRL uptake) or 24 h (lower left and middle panels for lipid accumulation). Lower right panel: representative FACS examples of lipid accumulation in THP-1 monocytes treated with TGRL for 48 h in the presence or the absence of exogenous LPL. D: Representative FACS examples and quantification of effects of TGRL treatment on THP-1 CD36 expression and uptake of OxLDL. THP-1 monocytes were treated with TGRL for 48 h and then, after being washed with PBS to remove TGRL, incubated with DiI-OxLDL in the presence or the absence of a CD36 mAb for an additional 4 h. THP-1 expression of CD36 was examined after incubation with TGRL and before incubation with DiI-OxLDL, and THP-1 uptake of DiI-OxLDL was examined after incubation with DiI-OxLDL. E: Effects of TGRL on THP-1 expression of IL-6, MCP-1, and IL-1β examined by quantitative RT-PCR. All representative examples were from ≥3 independent experiments with similar results. Data are shown as mean ± SEM.

Article Snippet: Monocyte phenotypes were analyzed by FACS using monoclonal antibodies (mAbs) to mouse antigens, including CD115 (PE, AFS98, ThermoFisher Scientific, Waltham, MA; or BV421, AFS98, BioLegend, San Diego, CA), CD11c (PerCP-Cy5.5, N418, ThermoFisher Scientific; or APC or FITC, N418, BioLegend), CD36 (FITC, MF3, Bio-Rad Laboratories, Hercules, CA), CD204 (FITC, 2F8, Bio-Rad Laboratories), Ly-6C (APC, HK1.4, ThermoFisher Scientific), CX3CR1 (FITC, SA011F11, BioLegend), MHCII (I-A/I-E; PE, M5/114, BD Biosciences, San Jose, CA), CD43 (PE, S11, BioLegend), TREML4 (PE, 16E5, BioLegend), XCR1 (PE, ZET, BioLegend), tumor necrosis factor α (TNFα; PE, MP6-XT22, ThermoFisher Scientific), and interleukin-1β (IL-1β; PE, NJTEN3, ThermoFisher Scientific), with appropriate isotype-negative controls.

Techniques: In Vivo, Ex Vivo, Isolation, Labeling, Injection, Staining, Incubation, Inhibition, Expressing, Quantitative RT-PCR

Journal: Journal of Lipid Research

Article Title: Foamy monocytes and atherogenesis in mice with combined hyperlipidemia and effects of antisense knockdown of apoCIII

doi: 10.1016/j.jlr.2025.100763

Figure Lengend Snippet: Effects of apoCIII ASO treatment on plasma lipids, monocyte phenotypes, and atherosclerosis in Ldlr −/− apoCIIItg mice. Ldlr −/− ApoCIIItg mice fed WD were treated with a GalNac-conjugated ASO against human apoCIII or a GalNac-conjugated CO (and Ldlr −/− mice were treated with CO) at 10 mg/kg body weight weekly for 12 weeks. A: Plasma total TG and cholesterol levels (n = 14–19 mice/group from three independent experiments with 4–7 mice/group in each experiment). B: Representative FACS examples showing SSC and CD11c of CD36 + and CD36 – monocytes (treatment for 4 weeks) and quantification of changes in proportions, SSC, and CD11c MFI of CD36 + monocytes with treatment (n = 11–14 mice/group). C: Representative photographs and quantification of en face Oil Red O staining of whole aorta. D: Representative photographs and quantification of Oil Red O staining in aortic sinus. E: Representative photographs and quantification of CD11c staining in aortic sinus lesions. Data are shown as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 compared to Ldlr −/− mice with CO (A and B); ## P < 0.01, ### P < 0.001 compared to Ldlr −/− ApoCIIItg mice with CO (A and B).

Article Snippet: Monocyte phenotypes were analyzed by FACS using monoclonal antibodies (mAbs) to mouse antigens, including CD115 (PE, AFS98, ThermoFisher Scientific, Waltham, MA; or BV421, AFS98, BioLegend, San Diego, CA), CD11c (PerCP-Cy5.5, N418, ThermoFisher Scientific; or APC or FITC, N418, BioLegend), CD36 (FITC, MF3, Bio-Rad Laboratories, Hercules, CA), CD204 (FITC, 2F8, Bio-Rad Laboratories), Ly-6C (APC, HK1.4, ThermoFisher Scientific), CX3CR1 (FITC, SA011F11, BioLegend), MHCII (I-A/I-E; PE, M5/114, BD Biosciences, San Jose, CA), CD43 (PE, S11, BioLegend), TREML4 (PE, 16E5, BioLegend), XCR1 (PE, ZET, BioLegend), tumor necrosis factor α (TNFα; PE, MP6-XT22, ThermoFisher Scientific), and interleukin-1β (IL-1β; PE, NJTEN3, ThermoFisher Scientific), with appropriate isotype-negative controls.

Techniques: Clinical Proteomics, Staining