Fig. 5.

Functional verification of the targeting siRNA tLyp-1 exosomes in silencing SOX2 gene of human NSCLC cells. (A) Designed 3 kinds of sequences for siRNA according to human SOX2 siRNAs. (B) mRNA expression of SOX2 in A549 cells after treated with lipo3000 carrying siRNA, natural exosomes carrying siRNA or targeting tLyp-1 exosomes carrying siRNA. Real-time qRT-PCR analysis was performed on A549 cells that were treated with lipo3000, natural exosomes or targeting tLyp-1 exosomes which carried 3 kinds of siRNAs, respectively. The results demonstrate that siR2 has the highest silence efficiency. P < 0.05; (a): vs natural exosomes; (b): vs. lipo3000. Data are presented as mean ± SD (n = 3). (C) Images of human NSCLC A549 cells under fluorescence microscope. (a): image of transfected A549 cells with siR2 lipo3000 under fluorescent microscope in bright field; (b): image of transfected A549 cells with the targeting siR2 tLyp-1 exosomes under fluorescent microscope in bright field; (c): image of transfected A549 cells with siR2 lipo3000 under fluorescent microscope in dark field; d, image of transfected A549 cells with targeting tLyp-1 exosomes under fluorescent microscope in dark field. siRNA (siR2) was labeled with Cy3 and blue was set as an indicator of transfection efficiency by fluorescence microscopy. The results demonstrate a successful transfection in A549 cells using the targeting siR2tLyp-1 exosomes.