American Society of Hematology

Figure 4.

$FPN-GFP expressed in primary human M-CSF–derived macrophages is rapidly removed from phagosomes. (A) Primary human macrophages, transduced with a lentivirus-producing FPN-GFP, are shown after having been exposed to IgG opsonized for only 4 minutes (top row) or 14 minutes (middle and bottom rows). Shown at the 14-minute time point are macrophages from 2 independent PBMC donors. Extracellular beads (in blue) at 4 and 14 minutes were marked with an anti-human AlexaFluor-647-conjugated secondary antibody. The macrophage plasmalemma was labeled with TRITC-conjugated WGA (in red). The white arrows point to FPN-GFP–positive phagosomes, whereas the arrowheads point to FPN-GFP–negative phagosomes that are present at the 14-minute timepoint. The representative micrographs were taken of fixed samples using laser scanning confocal microscopy. Scale bars, 10 μm. (B) The fraction of FPN-GFP–positive phagosomes in primary human M-CSF–derived macrophages expressing FPN-GFP at 4 and 14 minutes postphagocytosis is shown. These data derive from 2 independent experiments using 2 independent blood donors, and the graph represents the mean ± SD. Statistical significance was determined using an unpaired Student t test with a Welch’s correction. ****P < .0001.$

FPN-GFP expressed in primary human M-CSF–derived macrophages is rapidly removed from phagosomes. (A) Primary human macrophages, transduced with a lentivirus-producing FPN-GFP, are shown after having been exposed to IgG opsonized for only 4 minutes (top row) or 14 minutes (middle and bottom rows). Shown at the 14-minute time point are macrophages from 2 independent PBMC donors. Extracellular beads (in blue) at 4 and 14 minutes were marked with an anti-human AlexaFluor-647-conjugated secondary antibody. The macrophage plasmalemma was labeled with TRITC-conjugated WGA (in red). The white arrows point to FPN-GFP–positive phagosomes, whereas the arrowheads point to FPN-GFP–negative phagosomes that are present at the 14-minute timepoint. The representative micrographs were taken of fixed samples using laser scanning confocal microscopy. Scale bars, 10 μm. (B) The fraction of FPN-GFP–positive phagosomes in primary human M-CSF–derived macrophages expressing FPN-GFP at 4 and 14 minutes postphagocytosis is shown. These data derive from 2 independent experiments using 2 independent blood donors, and the graph represents the mean ± SD. Statistical significance was determined using an unpaired Student t test with a Welch’s correction. ****P < .0001.

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