Figure 9

Overexpression of FUS mutations and GFP-SNPH alters FUS-SNPH interactions in transfected neurons. (A) Representative images of primary cortical neurons transfected with HA-FUS^WT, HA-FUS^R514G or HA-FUS^ΔNLS (green). eGFP-SNPH is shown for each transfected cell which allowed PLA (red) between HA and eGFP to assess interactions within the cell body for each neuron analysed. Nuclei were counterstained with DAPI. A zoomed in image of the soma for each condition has been provided to show the change in FUS-SNPH interactions, Scale bar = 50 μm (B) Quantification of PLA interactions within soma for HA and eGFP. There is a significant decrease in the FUS-SNPH interaction in HA-FUS^R514G transfected cells compared to HA-FUS^WT (p < 0.01) and a non-significant decrease in the FUS-SNPH interaction in the HA-FUS ^ΔNLS compared to HA-FUS^WT (p = 0.6277). Statistical analysis was performed using a One-Way ANOVA with a post-hoc Tukey’s multiple comparisons test; error bars are ± SEM. N = five cells were analysed from three different independent replicates. Single antibody controls were performed for the antibody which shows a mainly diffuse pattern. *p < 0.05, **p < 0.01, ***p < 0.001 ****p < 0.0001.