Figure 1 | Scientific Reports

Figure 1

From: The neurorepellent, Slit2, prevents macrophage lipid loading by inhibiting CD36-dependent binding and internalization of oxidized low-density lipoprotein

Figure 1

Human and murine macrophages express the Slit2 receptor, Robo-1. (a), Human peripheral blood mononuclear cells (PBMC) were isolated and incubated with M-CSF or GM-CSF to generate M-CSF- or GM-CSF-induced macrophages, respectively. RT-PCR was performed using specific primers for Robo-1 and GAPDH. (b), Lysates were collected from M-CSF- and GM-CSF-induced human macrophages and immunoblotting was performed using antibodies specifically detecting Robo-1 or ß-actin. (c), M-CSF-induced human macrophages (top) and DLD-1 cells (bottom) were first incubated with viability dye followed by AF-594-conjugated secondary Ab (Control, blue peak), or anti-Robo-1 Ab and AF-594-conjugated secondary Ab (Sample, red peak). Data from live, single cells was acquired using an LSRII flow cytometer (BD Biosciences-US) with FACSDiva software (BD Biosciences-US) and analyzed with FlowJo v10 (BD Biosciences-US). (d), M-CSF- and GM-CSF-induced human macrophages, murine BMDM and RAW264.7 macrophages were fixed and incubated with anti-Robo-1 Ab, followed by Dylight549-conjugated secondary Ab (red) and AF-647-conjugated wheat germ agglutinin (pseudocoloured green). Cells were imaged using a spinning disk confocal microscope at 63× magnification. Scale bar, 10 μm.

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