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. 2011 Jun 7;39(17):7740–7749. doi: 10.1093/nar/gkr456

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — Single molecule FISH combined with immunofluorescence shows mRNA co-localization with RISC and P body proteins when miRNA is expressed. (A–D) smFISH-IF co-staining of protein antibody (green, Cy-3) and luciferase reporter mRNA (red, Cy-5) at various times after doxycycline induction of mRNA. Blue = DAPI. White arrows point to examples of co-localization. Yellow arrows point to co-localization shown at 5× size in the inset of the same picture. (A) Hedls, 0 h. (B) Hedls, 24 h. (C) Argonaute, 0 h. (D) Argonaute2, 24 h. (E) Luciferase reporter mRNA levels are reduced when miRNA is expressed. Average number of cytoplasmic mRNA foci per cell 0, 6, 12 and 24 h after doxycycline induction of miRNA. N = 52 cells for 0 h, 48 cells for 6 h, 51 cells for 12 h and 50 cells for 24 h. Error bars = 95% confidence interval for means.