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. 2022 Aug 19;13:975291. doi: 10.3389/fphar.2022.975291

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Copyright © 2022 Liu, Cao, Liu, Gao, Du, Wang, Cheng, Zhu, Jia, Pan, Zhang, Jiang, He, Xu, Zhang, An, Guo and Gu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Cellular uptake of T7-siYY1-exo in vitro and BBB/BBTB model. (A) The delivery efficiency of Free siYY1 and T7-siYY1-exo in LN229 cells. Cy3-labeled siYY1 and loaded DID-labeled exosomes (5 μg/ml) were used to treat LN229 cells. After 24h incubation, the cytoskeleton was stained with phalloidin and the cell nuclei were stained with DAPI. The fluorescent was photographed under a confocal laser scanning microscope using Nikon NIS-Elements software (Nikon, Tokyo, Japan). (B) The knockdown efficiency of YY1 was detected by Western blot. (C) Schematic diagram of the BBB model in-vitro. (D) Representative immunofluorescence images showing T7-siYY1-exo uptake into LN229 cells after passing through a bEnd.3 monolayer. Unmod-exo/T7-siYY1-exo (5 μg/ml) was added to upper compartment. The fluorescent was photographed under a confocal laser scanning microscope using Nikon NIS-Elements software (Nikon, Tokyo, Japan). (E) The positive rates of DiD and Cy3 were detected by flow cytometry. (F,G) The penetrating and tumor targeting efficacy of T7-siYY1-exo evaluated through BBTB/LN229 tumor spheroids co-culture model. The treatment concentration of unmod-exo/T7-siYY1-exo was 5 μg/ml. The fluorescent was photographed under a confocal laser scanning microscope using Nikon NIS-Elements software (Nikon, Tokyo, Japan).