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Fig. S1. Aggregated human γ-globulin (AHG) binding to CD4+ T cells from peripheral blood mononuclear cells (PBMC) of systemic lupus erythematosus (SLE) patients. Gates were first drawn to select CD4+lymphocytes (a). Subsequently, CD4+ T cells were analysed for AHG binding in the CD25 and CD25+ populations (b).

Fig. S2. Human CD4+ T cells stained with anti-FcγRIIIA/B antibody (a), anti-FcγRIIIB antibody (b) and overlay (c). Arrows mark the receptor protein in the cells. Images captured at ×630 magnification.

Fig. S3. Membrane rafts (MR) (green) stained using cholera toxin-B (CTB)−fluorescein isothiocyanate (FITC) and anti-FcγRIIIB (red). Aggregation of MR is observed with association of FcγRIIIB. Nuclei stained with 4',6-diamidino-2-phenylindole (DAPI). Arrows point to aggregated MR and receptor.

Fig. S4. Human naive CD4+ T cells show aggregation of membrane rafts (MRs) (green) underneath the C5b-9 (red). C5b-9 assembled with purified complement proteins C5b-6, C7, C8 and AlexaFluor® 594 (red)-labelled C9. C8 omission during assembly prevented the assembly of membrane attack complex (MAC) and MR aggregation (not shown).

Fig. S5. CD4+ T cells treated with immune complexes (ICs) and terminal complement complex (TCC) show aggregation of membrane rafts (MRs) (green) and associate with FcγRIIIA/B (red). Cells stained for MR (green) and FcγRIIIA/B (red). Images captured in phase contrast. MR and FcγRIIIA/B (a) and with overlay of cell images (b). Nuclei stained with 4',6-diamidino-2-phenylindole (DAPI).

Fig. S6. Immunoprecipitates (lanes 1 and 3) and immunoblots (lanes 2 and 4) obtained using anti-FcγRIIIA/B. CD4+ T cells (lanes 1 and 2) and Jurkat cells (lanes 3 and 4). Silver staining of immunoprecipitates, CD4+ T cells (lane 1) and Jurkat cells (lane 3). Immunoprecipitates analysed by Western blotting using anti-FcγRIIIA/B, lane 2 (CD4+ T cells) and lane 4 (Jurkat cells). At 29 kD a broad band with another band at 35 kD was observed in CD4+ T cells and a single band at 29 kD in Jurkat cells.

Fig. S7. Immunoprecipitates obtained using anti-FcγRIIIA/B monoclonal antibodies. Proteins stained using silver (left) and blots stained with Coomassie Brilliant Blue R250. Untreated cells (lane 1), terminal complement complex (TCC) (lane 2), TCC and immune complexes (ICs) (lane 3) and IC-treated cells (lane 4). Arrow point to a protein migrating at approximately 72 kD (Syk).

Fig. S8. Inhibition of aggregated human γ-globulin (AHG) binding to CD4+ T cells by anti-FcγRIIIA/B monoclonal antibody. 1 × 106 cells treated with AHG-AlexaFluor®488 (5 µg), control cells treated with isotype antibody (10 µg, left panel) and monoclonal anti-FcγRIIIA/B (10 µg, right panel).

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