Figure 6

5-FUrd alleviates cross-seeding of SOD1 by TDP-43.A, immunocytochemistry of HEK293FT cells 48 h post co-transfection with TDP-43^ΔNLS (red) and SOD^G85R-GFP reporter protein (green) in the presence of vehicle (top) or 1 μM 5-FUrd (bottom), demonstrating a reduction in SOD1 aggregates with 5-FUrd. Scale bar represents 50 μm. B, time-lapse live-cell microscopy of SOD^G85R-GFP cross-seeding by TDP-43^ΔNLS in the presence of 1 or 5 μM 5-FUrd shows a reduction in aggregate accumulation proportionate to drug concentration. Images were acquired every 30 min for 30 h. C and D, flow cytometry of saponin-treated cells co-transfected with the reporter protein with either TDP-43^ΔNLS (C) or WT TDP-43 (D) showing a reduced number of SOD1^G85R-GFP aggregate containing cells when treated with 5-FUrd. For each experiment, error bars represent SEM of 8 to 13 biological replicates. Statistical significance was determined using one-way ANOVA and Dunnett’s test for multiple comparisons (∗∗p < 0.01; ∗∗∗∗p < 0.0001). SOD1, superoxide dismutase; TDP, TAR DNA-binding protein.