Fig. 5 Effector function of MIH5 isotype variants.

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Fig. 5 Effector function of MIH5 isotype variants. Effector function of MIH5 isotype variants. ADCC in vitro assay (A) to compare effector function of MIH5 isotype variants. MC38 cells were 51Cr-labeled, opsonized with different MIH5 isotype variants, and exposed to whole blood from C57BL/6 mice for 4 hours. Specific lysis was quantified by measuring 51Cr release. n = 3, mean ± SEM. *P < 0.05. (B) Experimental setup of in vivo depletion assay. Splenic B cells labeled with Violet and Red tracer dye were used as target cells for in vivo depletion. The violet B cells were opsonized with mIgG2asilent variant, while the red B cells were opsonized with mIgG2a, mIgG2b, or mIgA variant. Subsequently, B cells were mixed 1:1 (top plot and diagram) and injected intravenously into C57BL/6 mice. Twenty-four hours later, the spleens were isolated, and the ratios between violet and red B cells were determined via flow cytometry (bottom plot and diagram). The ratios before injection and after spleen isolation were used to quantify the isotype-specific depletion of the target cells. (C) Representative experiment indicating mIgG2a, mIgG2b, and mIgA specific depletion of target cells with mIgG2asilent opsonized cells as reference population. n = 3, mean ± SEM. *P < 0.05. Johan M. S. van der Schoot et al. Sci Adv 2019;5:eaaw1822 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).