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. 2015 Jan 5;5:1. doi: 10.1186/2045-3701-5-1

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© Keppeke et al.; licensee BioMed Central. 2015

This article is published under license to BioMed Central Ltd. 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 use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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IMPDH2-GFP-tagged RR induced in live COS-7 cells are detected in stable and stationary state. COS-7 cells were transfected with IMPDH2-GFP plasmid and maintained in medium containing 1 mM ribavirin for 18 h. (A) Sequential pictures were captured for >10 min and representative images are shown at 2 min intervals. Similar to the antibody labeling experiment, changes in shape and size of RR were not observed. (B) Transfected cells were fixed and labeled by IIF with rabbit anti-IMPDH2 antibody (red). Cells with a high level of transfection (arrows, left panel) do not show RR (arrows). Cells with low transfection levels show RR (arrowheads). Of 336 cells counted, 16% show IMPDH2-GFP RR; transfection efficiency was 37%. Cells not transfected by IMPDH2-GFP showed RR labeled by anti-IMPDH2 antibody (double-arrows). All transfected IMPDH2-GFP-tagged RR (green, arrowheads) were labeled by rabbit anti-IMPDH2 antibody (red). Nuclei were counterstained with DAPI (blue). Data represent three in (A) and two in (B) independent experiments. Bars: 10 μm.