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. 2023 Nov 16;14:1237016. doi: 10.3389/fimmu.2023.1237016

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Copyright © 2023 Krishnan, Sendra, Patel, Lad, Greene, Smyth, Gallaher, Herron, Scott, Genead and Tolentino

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Binding assay of PolySia-NPs to Siglecs and macrophages. The NPs containing PEG binding to the Siglecs were detected using anti-PEG biotin/horseradish peroxidase (HRP) system according to manufacturer’s ELISA protocol (A). PolySia-NPs (square shape) and a blank-NPs control (round shape) at different concentrations (0.2−10mg/mL) were assessed for binging to Siglec-7 (red), -9 (blue), -11 (green) and Siglec-E (black) (A-i) and Siglec-3, -5 and -8 (A-ii). Mean ± SEM. LPS activated THP-1 cells (B), M0 macrophages (C), M1 macrophages (D) or M2 macrophages (E) were incubated with FITC-conjugated PolySia-NPs or FITC-conjugated Blank-NPs controls(green). Macrophages were fixed and stained for marker Iba-1 (blue). The representative image shows macrophages imaged for FITC conjugated PolySia-NPs or blank-NPs in green (1st column), IBA1+ macrophages in blue (2nd column) and merged colors with zoomed image (3rd column). NPs: nanoparticles. 50μm scale bar. Magnification: 100x.