Figure 8.

Heat shock increased the cytoplasmic localization of the H517Q mutant and triggered accumulation of R521G, R495X and G515X into stress granules in vivo. Confocal slices of zebrafish spinal cord from 25 hpf embryos incubated at 42.5°C for 45 min followed by immediate fixation and double immunostaining with anti-GFP (green) and anti-TIAR (red). For a group of spinal cord cells from embryos expressing the H517Q mutant, the GFP redistributed to the cytoplasm in a granular pattern without incorporating prominently into stress granules, whereas this occurred only rarely for cells expressing GFP-WT FUS. For embryos expressing the R521G, R495X and G515X mutants, the GFP assembled into aggregates that co-localized with anti-TIAR staining. Nuclei were stained with Draq5. Left panel obtained at 100×, right panels obtained using 3.44× optical zoom. Scale bars, 10 µm.