Fig. 4: Multimerization of minibinders leads to NF-κB activation.

a Multivalent minibinders were generated by fusing two or more tandem repeats of the minibinder together using 12, 16, or 40-residue (GlySer) linkers. Antiparallel 8.6 dimers were generated by fusing minibinder 8.6 to the C terminus of an antiparallel coiled-coil derived from myosin 10. b TLR3 is expressed on the cell surface and in the endosome of TLR3hi cells, which express an NF-κB-linked GFP reporter. GFP levels are measured with flow cytometry. The schematic was rendered using BioRender.com. c Left, Histograms showing GFP signal in stimulated TLR3hi cells. The identities and concentrations of the stimuli are provided on the right and left sides of each panel, respectively. Right, mean fluorescence intensity (MFI) values from assay duplicates.