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. 2019 Oct 15;18:727–738. doi: 10.1016/j.omtn.2019.10.005

Figure 5.

Figure 5

CD63 Is the Binding Target of Aptamer LL4A

(A) Binding capacity of LL4A to Mel28-PLX cells pretreatment Proteinase K or trypsin was measured by flow cytometry. (B) Colloidal silver-stained SDS-PAGE used to analyze the LL4A-assisted target purification. (C) The direct interaction between LL4A and CD63 in Mel28-PLX cells was detected using an aptamer pull-down assay, and the CD63 protein was examined by western blotting. (D) The co-localization of Cy5-labeled LL4A and anti-CD63 in Mel28-PLX cells was investigated by confocal microscopy imaging. Top panels: scale bars, 25 μm; bottom panels: scale bars, 10 μm. (E) The knockdown efficiency of CD63 was determined by western blotting. (F) Binding capacity of LL4A in CD63 knockdown Mel28-PLX cells and control cells was measured by flow cytometry. (G) The overexpression efficiency of CD63 in A375-PLX cells was determined by western blotting. (H) Binding capacity of LL4A in CD63 overexpressed A375-PLX cells and control cells was measured by flow cytometry. (I) Dissociation constant of LL4A for purified His-CD63 recombinant protein was determined by ELONA.