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. 2020 Oct 9;11:5091. doi: 10.1038/s41467-020-18907-6

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© The Author(s) 2020

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Depiction of molecular features built into the new Siglec-Fc construct. b Coomassie-stained SDS-PAGE gel of all Siglecs purified through a three-step Nickel, ST, and SEC column purification strategy. Results are representative of two independent sets of purifications. c Different amounts of purified Siglec-1-Fc was incubated with U937 cells in either a two-step or a pre-complexed approach with a constant amount of Strep-Tactin-AF647. d Flow cytometry results for probing Siglec-1-Fc WT or Siglec-1-Fc R116A with U937 cells using the optimized pre-complexed conditions. e Flow cytometry results of probing CD33-Fc WT with U937 cells in a two-step assay or a pre-complexing assay with αhIgG1-AF647 or Strep-Tactin-AF647. Error bars represent ± standard deviation of three replicates. Statistical significance calculated based on a two-tailed unpaired Student’s t-test.