Fig. 4
From: Locally anchoring enzymes to tissues via extracellular glycan recognition
Extracellular signaling activity of monomeric G3 fusion proteins and trimeric nanoassemblies. a Micrographs demonstrating NL-G3 and NL-TT-G3 bioluminescence on Jurkat T cells using a blue fluorescence filter. b Micrographs demonstrating GFP-G3 and GFP-TT-G3 fluorescence on Jurkat T cells using a green fluorescence filter. c Amount of NL or GFP bound to Jurkat T cells after 4 h incubation with NL-G3, NL-TT-G3, GFP-G3, or GFP-TT-G3. d Bright-field micrographs of Jurkat T cells incubated with PBS (untreated, negative control), WT-G3 (positive control), NL-G3, or NL-TT-G3 for 4 h. e Percentage of metabolic activity of Jurkat T cells incubated with WT-G3 (positive control), NL-G3, or NL-TT-G3 for 4 h. f hIL-2 produced by Jurkat T cells after incubation with PBS (negative control), WT-G3 (positive control), NL-G3, or NL-TT-G3 for 24 h. Lactose was added as an inhibitor of G3 binding to Jurkat T cells. g Asialofetuin (ASF) (7 µM) precipitation with various quantities of WT-G3, NL-G3, or NL-TT-G3 as measured by light scattering and absorbance at 420 nm. N = 4, mean ± s.d., ***p < 0.001, Student’s t test for c. N = 4, mean ± s.d., n.s. is no significant differences, ****p < 0.0001, ANOVA with Tukey’s post hoc for e and f. N = 3, mean ± s.d. for g. Scale bar = 25 µm for a, b. Scale bar = 50 µm for d. Data points at or above baseline signal are shown as open circles in c, e, f. Data for WT-G3 appear as gray bars/squares/traces, for monomeric G3 fusion proteins as blue bars/circles/traces, and for trimeric nanoassemblies as red bars/triangles/traces
