Figure 4
From: Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells
SOD1-S32 is resistant to seeded aggregation in cells. (A) Images from time-lapse microscopy of seeded aggregation. (B) Algorithm counting of aggregates in living cells from 24 to 48 h post co-transfection with G85R-GFP reporter protein and indicated non-tagged construct. Images were acquired every 30 minutes. Inset indicates the rate of inclusion formation per hour during the inclusion growth phase. (C) Co-transfection of G85R-W32-GFP and G85R-W32S-GFP with G85R-tdTomato shows clear intense GFP fluorescence co-localized with tdTomato fluorescence for G85R-W32-GFP but not G85R-W32S-GFP. (D) Analysis of the GFP fluorescence intensity that overlaps with the G85R-TdTomato inclusions shows that G85R-W32S-GFP intensity is significantly lower than that of G85R-W32-GFP. Error bars on bar charts and line plots indicate SEM of at least 3 biological replicates. For panel D box-whisker plots, centre line-mean; box limits-upper and lower quartile; whiskers-0 and 100% range. Significance was determined via students t-test (***p < 0.001). Scale bar, 50 µm (A); 10 µm (C).
