Figure 4:

GFP silencing in MDA-MB-231/GFP cells treated with functionalized anti-GFP NANPs loaded into exosomes and inhibition of NF-kB function in HEK-Blue™ hTLR3 and THP1-Dual™ cells. (A) Fluorescent microscope images of MDA-MB-231-GFPs taken 72 hours post treatment with exosomes and exosomes loaded with either NANPs. (B) Flow cytometry data of the mean GFP fluorescence and RT-qPCR analysis of the relative GFP expression of MDA-MB-231-GFP cells after 72 hours of incubation with NANPs loaded exosomes. Statistical significance for the samples compared to the negative control is denoted by * (*** p<0.001). Statistical significance of Exo/FS, Exo/FA compared to Exo/FS+Exo/FA is denoted by # (### p<0.001). (C) Schematic demonstration of the TLR3 signaling that activates the NF-kB pathway and the assembly of the NF-kB decoys (upon re-association of the RNA/DNA fibers) that inhibits the nuclear translocation of the activated NF-kB. Poly (I:C) is used to activate the TLR3 ligand, the TLR3 signal resulted in the activation of NF-kB and subsequently NF-kB entered the nucleus, where it will bind to specific sequences of DNA to promote the downstream transcription and translation of secreted embryonic alkaline phosphatase (SEAP). The reporter cell line HEK-Blue™ hTLR3 was transfected with fibers and stimulated with Poly (I:C). The cells were incubated for 24 hours, and the levels of NF-kB-dependent SEAP were measured in the supernatants. (D) Schematic demonstration of the TLR1/2 signaling that activates the NF-kB pathway that can be inhibited with the NF-kB decoys assembled upon re-association of RNA/DNA fibers (F(S) and F(A)).