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. 2011 Jul 27;39(20):8938–8951. doi: 10.1093/nar/gkr608

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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 8. — MBNL1 protein binds CAG and CUG repeats in vitro and co-localizes with repeat region of mutant HTT and ATXN3 transcripts in nuclear inclusions. (A) Combined FISH/IF assay using Cy3-labeled CAG probe in DM1 cells and CTG probe in normal, HD and SCA3 cells as well as rabbit polyclonal MBNL1 primary antibody (A2764) detected with secondary Cy2-labeled anti-rabbit antibody. Merged images show the colocalization of the MBNL1 protein with nuclear CAG repeat RNA inclusions in HD and SCA3 cells. Such colocalization is also demonstrated in DM1 cells with CUG RNA foci. In normal fibroblasts, MBNL1 is detected throughout the cell. DAPI staining (blue), MBNL1 (green), CAG and CUG nuclear RNA (red). (B) Results of filter binding assay to determine binding affinity of MBNL1 protein to 5′-end labeled transcripts of (CUG)₇, (CUG)₂₀, (CAG)₇, (CAG)₂₀, (AUG)₁₇, (UUA)₁₇ and (UGG)₁₇. Affinity of MBNL1 to CUG and CAG RNA is similar with only slight preference for the CUG repeats.