Fig. 5
From: Cell-based glycan arrays for probing glycan–glycan binding protein interactions
High-affinity Siglec-15 ligands bridge cell interactions. Lec2 CHO cells displaying high-affinity Siglec-15 ligands or unmodified Lec 2 CHO cells were stained with CTO and mixed with Siglec-15-expressing CHO cells that express extracellular Siglec-15 fused to GFP in the intracellular domain. The formation of cell clusters was determined by double staining and analyzed by flow cytometry and fluorescence microscopy. a Dot plot displaying GFP+ population (Siglec-15-expressing CHO cells) that are interacting with CTO-stained Lec2 CHO cells. b Percentage of GFP+ population (Siglec-15-expressing CHO cells) that interact with CTO-stained Lec2 CHO cells (n = 3, error bars are reported as SEM, significant difference compared with control was assessed using Student’s t-test (*P < 0.0001)). c Percentage of Siglec-15-expressing CHO cells (GFP+) that interact with CTO-stained Lec2 CHO cells by fluorescence microscopy (n = 3, error bars are reported as SEM, significant difference compared with control was assessed using Student’s t-test (*P < 0.0001)). d Fluorescence microscopy images of Siglec-15-expressing CHO cells (GFP+) and CTO-stained, unlabeled, Lec2 CHO cells (scale bar: 40 μm). e Fluorescence microscopy images of Siglec-15-expressing CHO cells (GFP+) interacting with CTO-stained Lec2 CHO cells labeled with α2-6-10 (scale bar: 40 μm)
