Figure 3.
Figure 3. Engagement of LFA-1 and CD16 induces more targeted degranulation at the IS than CD16 alone. (A) Fixed-cell imaging flow cytometry of YTS-CD16 cells conjugated with S2-IgG or S2-IC1-IgG cells. Quantitative analyses of area, mean fluorescence intensity (MFI), and total fluorescence intensity (area × MFI) of LysoTracker red (lytic granules; B) and CD107a (C) staining at the immunological synapse are shown as a feature of directed degranulation of YTS-CD16 cells. Data represent pooled results from three independent experiments; n > 100 cells/group. Error bars show ± SD. Live-cell confocal microscopy of YTS-CD16 cells transduced with a degranulation indicator LAMP1-pHluorin construct conjugated with S2-IgG or S2-IC1-IgG cells (D) or 10 µm polystyrene beads coated with anti-CD16 or anti-CD18 plus anti-CD16 antibody (E). Magenta, target cells; red, LysoTracker red (lytic granules); green, pHluorin (degranulation events). ****, P < 0.0001.

Engagement of LFA-1 and CD16 induces more targeted degranulation at the IS than CD16 alone. (A) Fixed-cell imaging flow cytometry of YTS-CD16 cells conjugated with S2-IgG or S2-IC1-IgG cells. Quantitative analyses of area, mean fluorescence intensity (MFI), and total fluorescence intensity (area × MFI) of LysoTracker red (lytic granules; B) and CD107a (C) staining at the immunological synapse are shown as a feature of directed degranulation of YTS-CD16 cells. Data represent pooled results from three independent experiments; n > 100 cells/group. Error bars show ± SD. Live-cell confocal microscopy of YTS-CD16 cells transduced with a degranulation indicator LAMP1-pHluorin construct conjugated with S2-IgG or S2-IC1-IgG cells (D) or 10 µm polystyrene beads coated with anti-CD16 or anti-CD18 plus anti-CD16 antibody (E). Magenta, target cells; red, LysoTracker red (lytic granules); green, pHluorin (degranulation events). ****, P < 0.0001.

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