Fig. 4: Validation of specificity using target-specific siRNA.

Cell lines were transfected with siRNA specific for each aptamer’s reported target (red) or a non-specific siRNA control (blue) and subsequently stained with a target-specific antibody or aptamer. Aptamer or antibody binding to untreated cells are shown in green and non-targeted controls (isotype antibody or the non-targeting aptamer C36) are shown in black. Untreated cells are indicated in gray. All assays were performed at 37 °C using our internalization and binding with blocking protocol except for those denoted with an asterisk. The asterisk denotes experiments performed in the absence of blocking agents (see text for details). The cell type and aptamer concentration used for each experiment were: A LNCaP cells: A9.min (100 nM), A10-3 (100 nM), and A10-3.2 (500 nM); B A431 cells: SGC8c (100 nM); C HeLa cells: Waz (100 nM) and C2.min (100 nM); D HeLa cells: E07 (500 nM) and CL4 (1000 nM); E SKBR3 cells: SE15-8-mini (1000 nM) and 2-2(t) (1000 nM); F MCF7 cells: EpDt3 (1000 nM); and G HeLa cells: GL21.T (1000 nM). Data are representative of two independent experimental replicates.