Figure 5.

Preparation of GFPpG oligomers. (A–C). The mixture GFPpG oligomers was purified by nickel affinity chromatography and analyzed using SDS-PAGE (CBB staining). (A) Non-boiled GFPpG preparation were analyzed by SDS PAGE and visualized with CBB/fluorescence. (B) The assembly of GFPpG oligomers was assessed by BN-PAGE under UV light. (C,D) Purity of isolated GFPpG oligomers monitored with BN-PAGE. (E) Evaluation of Fc-FGF1 and FGF1-Fc interaction with GFPpG oligomers with BLI. FGF1 variants were chemically immobilized on BLI sensors and incubated with each GFPpG oligomer. Association and dissociation profiles were measured. (F,G) Complex formation between FGF1 fusions with the Fc and GFPpG oligomers. Recombinant proteins: Fc-FGF1 and FGFG1-Fc and various GFPpG variants (from dimer to pentamer) were mixed and incubated for 5–10 min at RT. Then, proteins mixture was subjected to BN-PAGE analysis under UV light.