Figure 4 | Scientific Reports

Figure 4

From: Targeted delivery of TLR3 agonist to tumor cells with single chain antibody fragment-conjugated nanoparticles induces type I-interferon response and apoptosis

Figure 4

Selective delivery of scFv(h-AM1)-BAP-RICIA into PSCA-positive target cells causes NF-κB activation and subsequent release of IFN-β. (a) Expression levels of PSCA in HEK-BluehTLR3/PSCA reporter cell line and HT1376 cell line analyzed by flow cytometry analysis using scFv(h-AM1). (b) Confocal laser scanning microscopy reveals specific receptor-mediated internalization of FITC-mal19-PPI-buffered scFv(h-AM1)-BAP-RICIA into HEK-BluehTLR3/PSCA cells (see arrows) whereas no uptake is seen when using scFv(MR1.1)-BAP-conjugated control RICIA. (c) Treatment of HEK-BluehTLR3/PSCA with scFv(h-AM1)-BAP-RICIA resulted in the significant induction of secreted embryonic alkaline phosphatase (SEAP) activity assessed via optical density (OD) at 655 nm (mean ± SD; ***p < 0.001). (d) Inhibition of PSCA-mediated internalization of scFv(h-AM1)-BAP-RICIA after treatment with the inhibitors of endocytosis filipin III or chlorpromazine (mean ± SD; ***p < 0.001). (e) Targeted delivery of the TLR3 agonist Riboxxol formulated in scFv(h-AM1)-BAP-RICIA led to a dose-dependent release of IFN-β in HEK-BluehTLR3/PSCA (mean ± SD).

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