Fig. 2: Investigating Siglec–ganglioside interactions using optimized liposomes reveals interactions.
a Heatmap summarizing the binding interactions of each human Siglec (4 ≥ n ≥ 3 technical replicates). Color is representative of the log10(MFIAF647) of each GLL subtracted from the log10(MFIAF647) of the same GLL against UT CHO cells. †Denotes binding was measured after treatment of cells with neuraminidase. b Schematic of the enzyme-linked immunosorbent assay (ELISA) used to investigate Siglec–ganglioside interactions outside the context of a lipid bilayer. c ELISA results of Siglec-1, -6, and -7 binding to nine gangliosides (n = 4 technical replicates). d Schematic of the liposome over lectin assay (LOLA) where binding is read out on a fluorescence plate reader. e Results of the LOLA against a select number of GLLs (n = 5 technical replicates). f Schematic of the bead assay where binding is read out by flow cytometry. g Results of the bead assay against a select number of GLLs (n = 4 technical replicates). All results are represented as the mean ± one standard deviation of at least three technical replicates. The dotted line on the ELISA, LOLA, and bead assay results represents two standard deviations above the blank well background for the ELISA and a naked liposome for the LOLA and bead assay. For panels c, e, and g, a Brown–Forsythe and Welch one-way ANOVA was used. Not Significant (NS), P > 0.5; *0.05 > P ≥ 0.01; ** 0.01 > P ≥ 0.001; ***0.001 > P ≥ 0.0001; ****P < 0.0001.
