Recombinant proteins consisting of human IgG Fc and extracellular domains of human SIGLEC1, 2, 3, 5, 6, 7, 9, 10, 11, and mouse Siglec1, 2, E, F were purchased from R&D Systems (Minneapolis, MN). Recombinant mouse Siglec3-hIgGFc was purchased from Sino Biological, Inc. (Beijing, China) and anti-mouse TLR4 (MTS510) from Biolegend (San Diego, CA). Anti-human TLR4 and Anti-Siglec-E were from R&D. Anti-mouse CD11c, CD11b, CD4, CD8, B220 and Gr1 were purchased from eBioscience (San Diego, CA). Anti-Neu1 (H-300) and anti-Neu3 (M-50) antibodies and Horseradish perioxidase conjugated anti-mouse, anti-goat or anti-rabbit secondary-step reagents, as well as anti-TLRs antibodies that are cross-reactive for mouse and human TLR1 (H-90), TLR2 (A-9), TLR3 (M-300), TLR5 (M-300), TLR6 (N-18), TLR7 (N-20), TLR8 (H-114), TLR9 (H-100) and TLR10 (V-20) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA) and biotinylated for studies used in Figure 1A,B. Lipopolysaccharide (LPS, from E. coli 0111:B4) was from Sigma. E. coli (strain 25922) was obtained from ATCC (Manassas, VA). Neu5Ac2en and Neu5Gc2en were synthesized as described (Li et al., 2008 (link), 2011 (link)).
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