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. 2014 May 12;23(18):4945–4959. doi: 10.1093/hmg/ddu210

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© The Author 2014. Published by Oxford University Press.

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 reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — Interactions between the premutation CGG repeats and adaptation-promoting pathways. (A) Olfactory adaptation. 99CGG animals were crossed with mutants defining three pathways: ALG-2 is an miRNA-specific Argonaute, FBF-1 up-regulates EGL-4 translation, and MUT-7 is required for siRNA processing. alg-2 knockout decreased the effect of expanded CGG repeats, reducing adaptation defects in the double mutants. Conversely, the adaptation defects of 99CGG animals crossed with fbf-1 and mut-7 mutants were partially additive to defects in 99CGG animals alone. CI experiments were performed in at least triplicate, and error bars represent SEM. (B) Genotyping of animals with transgenes. 206- and 505-bp PCR fragments were amplified in animals carrying pAWC::FMR(CGG)₀::GFP and pAWC::FMR(CGG)₉₉::GFP plasmids. A few non-specific bands are indicated by ‘*’. (C) Expanded-repeat mRNA levels had no significant change after crossing with alg-2, fbf-1, and mut-7 knockout lines. Bars represent the fold change of FMR(CGG)₉₉ mRNA levels in pAWC::FMR(CGG)₉₉::GFP, or in either alg-2, fbf-1 or mut-7 double mutants, respectively, compared to the mRNA levels of a control pAWC::FMR(CGG)₀::GFP. The data were collected from four independent experiments.