Supplementary Figure 2: Expression of mbl-1::mCherry in C. elegans muscle cells increases expanded-CUG-transcript recruitment and nuclear-foci accumulation of mutant transcripts.
From: Identification of genes in toxicity pathways of trinucleotide-repeat RNA in C. elegans
Schematic drawing of the MBL-1::mCHERRY construct (A), and C. elegans body wall muscle cells (B). (C) MBL-1::mCHERRY exhibits a diffuse cellular distribution with nuclear accumulation. (D) C. elegans muscle cells confocal SM-FISH images of GFP RNA transcripts (white), DAPI stained nucleus (blue), merge of GFP RNA and nucleus images, and mCherry translational fusion protein. The imaged muscle cells correspond to animals expressing 123CUG repeats and 0CUG, in a mCHERRY (control) or MBL-1::mCHERRY backgrounds. Yellow arrows indicate expanded CUG nuclear foci. MBL-1::mCHERRY localizes to the nucleus. (E) Genetic mosaic analysis of GFP intensity shows that GFP fluorescence, from 123CUG mRNA transcripts, absent in cells expressing mbl-1::mCherry, relative to neighboring cells that fail to express mbl-1::mCherry. GFP fluorescence is not affected in the 0CUG control animals expressing mbl-1::mCherry. (F) Confocal SM-FISH images of GFP RNA transcripts (white), DAPI stained nucleus (blue) and merge of C. elegans muscle cells. The strains imaged were 123CUG and 0CUG in: empty vector control (ctrl) and mbl-1 gene inactivations. Yellow arrows indicate expanded CUG nuclear foci. Bar, 1 μm.
