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Figure S1: HLA I antibody binding to human aortic endothelial cells. Human aortic endothelial cells were stained with W6/32 (1 μg/mL). Antibody binding was detected by staining with goat antimouse-FITC secondary antibody. Representative flow cytometric histograms are shown, where the dashed line is unstained control and the dark line is HLA I antibody.

Figure S2: Dose-dependent response of P-selectin induction by HLA I crosslinking. (A) HAEC were treated with HLA I antibody at 1 μg/mL, 5 μg/mL or 10 μg/mL, or isotype control mIgG at 1 μg/mL for 30 min. (B) HAEC were treated with mIgG or HLA I antibody at 5 μg/mL for the indicated times. Cell surface P-selectin was stained with PE-conjugated anti-P-selectin. Graph shows (A) mean fold increase in P-selectin positive cells normalized to mIgG from multiple independent measurements (n=13) or (B) percentage of cells gated positive for P-selectin from one representative experiment out of three. (C) A representative histogram of P-selectin staining on unstimulated (gray fill) or HLA I antibody-activated (bold line) endothelial cells is shown.

Figure S3: Long-term exposure to HLA I antibodies does not alter endothelial expression of E-selectin. HAEC were treated with HLA I antibody or isotype control mIgG (5 μg/mL), or TNFα (10 pg/mL) for the indicated times. Cell surface E-selectin was stained with PE-conjugated anti-E-selectin. Results are expressed as median fluorescence intensity of E-selectin staining from one representative experiment out of two similar.

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