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. 2021 Jun 9;49(11):6456–6473. doi: 10.1093/nar/gkab484

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Optimization of RP-CONA method in the Opera HCS. (A) A diagram illustrating the interaction between FITC-pri-miR-7–1-CTL and mCherry-HuR. (B) Beads images of blank beads; blank beads incubated with cell lysates containing mCherry-HuR; FITC-pri-miR-7–1-CTL-beads incubated in lysates-free buffer; and FITC-pri-miR-7–1-CTL-beads after mCherry-HuR pulldown. (C andD) mCherry/FITC signals were reduced by untagged pri-miR-7–1-CTL. Cell lysates containing mCherry-HuR were treated with an increased concentration of untagged pri-miR-7–1-CTL before pull-down. (E and F) mCherry/FITC signals were reduced by a pri-miR-7–1/HuR inhibitor. Cell lysates containing mCherry-HuR were treated with DMSO or an increased concentration of OA before pull-down. All values were obtained from three technical repeats. Mean mCherry/FITC ring intensities and SD between triplicates are shown. Statistically significant differences compared to 0 μM untagged pri-miR-7–1 or DMSO were interpreted by SPSS independent sample t-test, ****P< 0.0001.