Supplementary Figure 1: Properties of MAB-hR3. | Nature Immunology

Supplementary Figure 1: Properties of MAB-hR3.

From: IL-1R3 blockade broadly attenuates the functions of six members of the IL-1 family, revealing their contribution to models of disease

Supplementary Figure 1

(a) Gating strategy for FACS analysis of MAB-hR3 and isotype binding to SK-MEL-30 and NIH-3T3 cell lines (Fig. 1b) (b) MAB-hR3 immunoprecipitate from A549 cell lysate. Gel electrophoresis of immunoprecipitate stained with collodial coomassie. The arrow at 80kDa indicate the primary location of IL-1R3 peptides as analyzed by MS. IL-1R3 peptides, presumably proteolytic breakdown products, were similar found at 60kDa (5 peptides) and 20kDa (3 peptides). (c) Viability of PBMCs using MAB-hR3 (20μg/mL or 1μg/mL) compared to media alone (percentage) (left y-axis) and viability of media alone as compared to 24hrs (right y-axis), using a WST-1 cell proliferation reagent at 24 hours, 3 days and 5 days, respectively. (d) IL-6 production in unstimulated PBMCs using MAB-hR3 (20μg/mL or 1μg/mL) alone. Assayed at 24 hours, 3 days and 5 days, respectively. Mean +SEM, data from 3 donors, all in triplicates.

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