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. 2014 Feb;20(2):260–266. doi: 10.1261/rna.041905.113

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© 2014 Sinnamon and Czaplinski; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.

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FIGURE 3. — Simultaneous detection of Actb and Actg mRNAs with FISH STIC probes. Two Actb and two Actg primary probes were synthesized, and corresponding secondary and tertiary probes were synthesized to label Actb mRNA with Cy3 and Actg mRNA with Cy5. Probes were hybridized to primary MEFs and imaged with epifluorescence microscope. (Top row) Normalized Cy3 images of cells after hybridization with complete FISH probes (A,B) or probes omitting one set of primaries as indicated above (C,D). (Middle row) Normalized Cy5 images of cells after hybridization with complete FISH probes (A′,B′) or probes omitting one set of primaries as indicated above (C′,D′). (Bottom row) Cy3 (red), Cy5 (green), and DAPI (blue) merged images of cells above it. (B,B′,B′′) Detailed images of the boxed regions of interest (ROI) in panels A, A′, and A′′, respectively. Scale bars, (columns A,C,D) 10 μm; (column B) 2 μm.