Moreover, after approximately five hours of chronic OptoGranule assembly, we observed a significant increase in immunopositivity using antibodies to phospho-TDP-43 and the ubiquitin-binding proteins SQSTM1 and VCP, illustrating further evolution of these structures (Figure 3g–i).Thus, not only does chronic OptoGranule assembly cause a loss of cell viability, but cell death is preceded by the evolution of OptoGranules into cytoplasmic inclusions that recapitulate features that are pathognomonic for ALS-FTD. This evidence concerns the gene VCP and amyotrophic lateral sclerosis.