Fig. 2. Cellular response to FolamiRs in vitro.

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Fig. 2. Cellular response to FolamiRs in vitro.

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Fig. 2. Cellular response to FolamiRs in vitro. Cellular response to FolamiRs in vitro. (A) Targeted silencing of miR-34a Renilla sensor using FolamiRs in MB-231 breast cancer cells. Data points were normalized to FolamiR-NC (negative control: scrambled miRNA) for each time point. (B) Proliferation of MB-231 cancer cells as a function of FolamiR treatment (50 nM). Data points were normalized to FolamiR-NC for each time point. Error bars: means ± SD. Each experiment corresponds to n = 3 with at least three technical replicates per treatment. (C) Dose response of MB-231 cells to FolamiR-34a. Renilla values were measured 96 hours after treatment. Data points were normalized to FolamiR-NC. Duplex miRNA represents unconjugated miR-34a. Error bars: means ± SD. Each experiment corresponds to n = 3 with at least four technical replicates per treatment. (D) Displacement of NIR-FolamiR-34a binding from MB-231 cells (50 nM, 4°C) with increasing concentrations of folate-glucosamine conjugate. Histograms represent overlaid flow cytometry data as a percentage of unstained and NIR-FolamiR-34a–stained cells. (E) FolamiR-34a competition assay in MB-231 breast cancer cells in vitro. MiR-34a Renilla sensor response to FolamiR-34a (50 nM, 96 hours) in the presence of increasing concentrations of folate-glucosamine conjugate. Data points were normalized to FolamiR-NC (negative control: scrambled miRNA) for each experimental condition. Experiment corresponds to n = 3 with at least three technical replicates per treatment. *P < 0.05, **P < 0.01, one-way ANOVA with post hoc Bonferroni correction. Esteban A. Orellana et al., Sci Transl Med 2017;9:eaam9327 Published by AAAS