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. 2019 Feb 26;2(2):e201800222. doi: 10.26508/lsa.201800222

Figure 3. FUS relocalises to nucleoli in response to TOP1-induced DNA breakage in neurons.

Figure 3.

(A) Endogenous FUS and B23 (nucleophosmin; nucleolar marker) were detected by indirect immunofluorescence in mouse cortical neurons following incubation with DMSO vehicle or 4 μM CPT for 45 min. Scale bar, 5 μm. (B) FUS-GFP was detected by direct fluorescence in human spinal motor neurons after treatment with either DMSO vehicle or 4 μM CPT for 1 h. Fibrillarin and MAP2 were detected by indirect immunofluorescence to stain nucleoli and neurons, respectively. Numbers are the mean percentage (±SD) of MAP-positive cells with FUS-GFP nucleolar localization in three independent experiments (50 cells per experiment). Scale bar, 10 μm. (C) FUS-GFP was detected by direct fluorescence in human spinal motor neurons after treatment with either DMSO vehicle or 4 μM CPT for 1 h. Choline acetyltransferase (ChAT) was detected by indirect immunofluorescence to confirm the identity of motor neurons. Numbers are the percentage of ChAT-positive cells with FUS-GFP nucleolar localization (n = 100 cells). Scale bar, 10 μm. Zoomed areas are shown (right).