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. 2019 Jan 8;10(1):17. doi: 10.1038/s41419-018-1267-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, b RIP (RNA Immuno Precipitation) assays performed in control (CTR) and miR-145-5p overexpressing (145-5p) MDA-MB-231 cells using antibodies directed to Ago2, Ago1, or IgG as negative control. mRNA abundance of Ago2 was evaluated by RT-qPCR and normalized over GAPDH mRNA; a representative experiment of a biological duplicate is shown. c Cytoplasmic extracts from MD-MB-231 cells overexpressing miR-145-5p and the relative control cells were loaded on 15–50% sucrose gradients and fractions measured by absorbance at 253 nm (left panel). Fraction density decreases from left to right; a representative profile out of two independent biological replicates is showed. Right panel: pooled fractions of RNA associated with polysomes, monosomes and free RNA were analyzed by RT-qPCR. The percentage of Ago2 mRNA enrichment, normalized over GAPDH mRNA is reported