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Blood, Vol. 111, Issue 8, 4245-4253, April 15, 2008
Targeting DCIR on human plasmacytoid dendritic cells results in antigen presentation and inhibits IFN- production
Blood Meyer-Wentrup et al.
111: 4245
Supplemental materials for: Meyer-Wentrup et al
Files in this Data Supplement:
- Figure S1. Murine monoclonal and goat polyclonal anti-DCIR antibodies do not cross-react with BDCA-2 (JPG, 66.5 KB)
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HEK293 cells transiently transfected with human BDCA-2 or DCIR DNA constructs were labeled with murine anti-BDCA-2, goat anti-DCIR, murine anti-DCIR or matching isotype control antibodies and analyzed by flow cytometry. The signals detected on transfected cells with the indicated antibodies (shaded areas) and matching isotype controls (dotted lines) are overlaid with the signals obtained from untransfected HEK293 cells stained with the antibodies (solid lines) or with isotype controls (dashed lines). The anti-DCIR antibodies detect DCIR, but do not cross-react with BDCA-2 and vice versa. FACS data are representative of two independent experiments.
- Figure S2. Dominant-negative dynamin inhibits DCIR endocytosis (JPG, 60.3 KB)
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The CHO cell line stably expressing DCIR was co-transfected with dynamin-WT or -DN constructs. Cells were incubated with anti-DCIR and secondary fluorochrome-conjugated antibodies. Surface-bound antibodies were stripped off as described in materials and methods and cells were analyzed by flow cytometry. Incubation at 37°C induced DCIR internalization in dynamin-WT co-transfected cells (left), while co-transfection with DN-dynamin inhibited DCIR internalization (middle). DCIR was well internalized into mock-transfected cells (right). The grey shaded areas indicate internalized DCIR at 37°C. The thin lines show DCIR internalization at 4°C, while the thick lines depict total surface DCIR expression. The data shown are representative of three independent experiments.
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