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. 2021 Jun 3;11:11676. doi: 10.1038/s41598-021-91112-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of DC-SIGN pre-exposure on anti-IgM-induced signaling in WSU-FSCCL cells. (a, b) Anti-IgM-induced iCa²⁺ flux in WSU-FSCCL cells following pre-exposure to DC-SIGN-Fc/HA. (a) Representative results for DC-SIGN-Fc (left) and DC-SIGN-HA (right) pre-treatment follow by addition (arrow) of the stimulation as stated and (b) summary of data mean for 6 (DC-SIGN-Fc) or 3 (DC-SIGN-HA) individual experiments. Graphs show mean (± SD) response with values for anti-IgM only cells set to 100%. Statistical significance of the indicated differences is shown (paired t-test). (c) DC-SIGN displacement after mannan incubation. FACS analysis of WSU-FSCCL cells left untreated (grey), incubated with DC-SIGN-Fc 30 min at 37 °C (blue) or incubated with DC-SIGN-Fc 30 min at 37 °C, washed and left 15 min with 40 μg/ml of Mannan at 37 °C (red). Cells were stained with APC Cy-7 F(ab’)₂ anti-hIgG Fc (α-IgG Fc) to detect DC-SIGN-Fc binding. (d, e) Effect of pre-treatment with DC-SIGN-Fc ± mannan for 15 or 30 min at 37 °C on iCa²⁺ flux. (d) Representative result and (e) summary of data mean for 3 individual experiments. Graphs show mean (± SD) response with values for anti-IgM only/no pretreatment cells set to 100%. Statistical significance of the indicated differences is shown (paired t-test).