Figure 4.

Apt69.T Aptamer Internalization and miRNA/anti-miR Delivery In Vitro

(A) Internalization of apt69.T on U266 (left) and H929 (right) (BCMA+) cells. Apt69.T (200 nM) was incubated for 15 min with cells and then cells were recovered following three washings with PBS (total) or with PBS 0.5 M NaCl (internalized) in order to remove aptamer on the cell surface. The quantities of total and internalized aptamer were evaluated by qRT-PCR. Error bars show the mean ± SEM values. (B) Schematic representation of the apt69.T-miR-137 conjugate (top) and apt69.T-anti-miR-222 (bottom). (C) U266 (BCMA+) or CCRF-CEM (BCMA−) cells were treated (400 nM) with indicated molecules. miR-137 level relative to cells treated with control aptamer (scraCL4) was evaluated by qRT-PCR following 48 h. Bars show the mean ± SD values. (D) U266 (BCMA+) and CCRF-CEM (BCMA−) cells were treated (400 nM) with indicated molecules. 72 h post-treatment, cell viability was measured by MTT assay. Error bars show the mean ± SD values. (E) NCI-H929 (BCMA+) or CCRF-CEM (BCMA−) cells were treated (400 nM) with indicated molecules. miR-222 level relative to cells treated with control conjugate (SCRA-222) was evaluated by qRT-PCR following 72 h. Error bars show the mean ± SD values.