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Figure 6 | Molecular Neurodegeneration

Figure 6

From: Amyotrophic lateral sclerosis-linked FUS/TLS alters stress granule assembly and dynamics

Figure 6

Methylation of mutant-FUS is not required for its assembly into stress granules. HeLa cells transfected with GFP-FUS R495X were pre-treated with the methyltransferase inhibitor adenosine-2,3 dialdehyde (AdOx) and then exposed to sodium arsenite to promote stress granule assembly. (A) GFP-FUS R495X was detected by western analysis only for immunoprecipitation (IP) reactions from transfected cells (GFP-FUS (+)). Anti-GFP was used for both IP and western analyses. The ASYM24 antibody revealed that GFP-FUS R495X was hypomethylated on arginine residues when cells were pre-treated with AdOx. (B) Confocal fluorescence images revealed a robust association of GFP-FUS R459X with sodium arsenite-induced (SA(+)) stress granules, both in the absence (left panels) and presence (right panels) of AdOx. Conversely, the ASYM24 signal within the same stress granule was dramatically attenuated when cells were pre-treated with AdOx. (C) Quantification of n=3 independent experiments from (B) further supports the conclusions above. Statistical significance was determined by a Student’s t-test (** P < 0.01). All error bars represent SEMs.

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