Picture from Lamitina, Todd (2022) MicroPubl Biol

Figure 1. Characterization of a C. elegans G4C2 repeat model. A) Description of G4C2 expression clones and their expression products. Characteristics of these expression clones are listed above the G4C2 repeat domain. B) In situ hybridization to detect G4C2 -containing mRNA in muscle cell nuclei. Arrowheads point to examples of RNA foci, which were only observed in muscle cell nuclei (dashed circle). Scale bar = 1 micron. C) Quantification of the percentage of animals with observable RNA foci. The number above each bar represents the number of animals examined. **** - p<0.0001, Fisher's exact test with Bonferroni correction. D) Canonical translation or RAN translation was detected using GFP expression. Note in the '+ATG' lines how GFP expression transitions from soluble to puncta as the number of repeats increase. This is consistent with the production of a (Gly-Ala)x-GFP fusion protein with increasing number of repeats, which is known to form aggregates in C. elegans (Rudich et al. 2017). Without a start ATG, GFP expression is not observed until repeats ≥33, which fits the definition of RAN translation (Zu et al. 2011). All images were taken using identical exposure times. Scale bar = 100 microns. E) Quantification of RAN translated GFP. Data were normalized to the mean of the 5 repeat measurements. Graph shows the mean ± S.D. with individual data points shown. **-p<0.01, ***-p<0.001, ****-p<0.0001, One-way ANOVA with Welch test.