All stainings with plant lectins (Vector Laboratories), Lectenz (Lectenz Bio) or Siglec-Fc chimeras (R&D systems) were performed in HBSS containing magnesium and calcium (Gibco) supplemented with 0.5% fatty-acid free BSA (Sigma). Siglec-Fc chimeras were pre-incubated at 1 μg/mL with anti-human IgG Fc (Biolegend, clone: HP6017) for 15 minutes at room temperature, after which they were added to the cells. Similarly, 1 μg/mL lectins and Lectenz reagents were pre-incubated with streptavidin-APC before addition to the cells. Macrophages in Fig. 4 were analyzed with a 14-color antibody panel (supplementary Table 2 ) on the Cytek Aurora and analyzed with FlowJo v10 and OMIQ. Fibroblast markers PD-L1, HLA-DR and α-SMA were measured on Cytek Aurora, other experiments were analyzed using the Fortessa™ X‐20 and analyzed with FlowJo v10 (list of antibodies in supplementary Table 2 ).
In indicated experiments, cells were treated at 37 °C for 30 minutes with neuraminidase from Arthrobacter ureafaciens (Roche Diagnostics, diluted 1:100). To assess the presence of sialylated structures in N-glycans, O-glycans, or glycolipids, cells were treated for 3 days with 10 μg/mL Kifunensine, 0.8 mM Benzyl-GalNAc, or 5 μM PPMP, respectively.
In indicated experiments, cells were treated at 37 °C for 30 minutes with neuraminidase from Arthrobacter ureafaciens (Roche Diagnostics, diluted 1:100). To assess the presence of sialylated structures in N-glycans, O-glycans, or glycolipids, cells were treated for 3 days with 10 μg/mL Kifunensine, 0.8 mM Benzyl-GalNAc, or 5 μM PPMP, respectively.