Amyotrophic lateral sclerosis (ALS) is one of the most common degenerative diseases of the motor neuron system. FUS/TLS and TDP-43, two strikingly similar RNA Binding Proteins, are dominant causes of both the fatal adult motor neuron disease amyotrophic lateral sclerosis (ALS) and frontal temporal degeneration (FTD). Stress granules (SGs) are granules of RNA and proteins formed in the cytosol of the cell under stressful conditions. Disease-linked mutations of FUS increase its propensity to aggregate and to form SGs by preventing nuclear translocation. FUS/TLS Stress Granules Assay Cell Line from Innoprot allows the identification of pathological stress granules formation inhibitors.

In normal conditions, FUS/TLS protein is predominantly localized in the nucleus. In the presence of an oxidative insult like Sodium arsenite, FUS/TLS increases its cytoplasmic localization forming Stress granules. This cell line allows the measurement of localization patterns to identify most promising test compounds.

To perform the assay, we incubate the cells with test compounds and Sodium Arsenite to induce the FUS/TLS aggregation. Finally, we quantify FUS/TLS-tGFP nuclear globs using the BD Pathway HCS Reader and Attovision Compartimentalization Software.

Applications:

• FUS/TLS Stress Granules Assay cell line allows the quantification of pathological FUS/TLS Stress Granules.
• This model permits to evaluate the FUS/TLS protein distribution in living cells studying the protein localization pattern in the space and time.
• This model provides a strategy to evaluate drugs without cell permeability.