Fig. 2.

FUS protein localization in mESC-derived MNs: a Multiple immunostaining analysis of FUS (red) and the MN marker Islet-1 (cyan) in FUS^WT, FUS^HOMO, and FUS^KO mixed cell populations cultured for 2 days after EB dissociation and replating. The higher GFP signal is mainly located in Islet1(+) cells. No significant FUS signal was found in FUS^KO cells. Nuclear staining with DAPI (4′,6-diamidino-2-phenylindole) is reported in blue. Scale bar: 20 μm. b Magnification of square inserts (A, B, C) reported in a showing FUS and Islet-1 double immunofluorescence (row I) or FUS staining combined with transmitted light (row II). c Representative line scan analysis of the FUS signal intensity in MNs indicated by asterisks in B (row II). A fluorescent intensity value along the black line drawn across the nucleus and cytoplasm (upper panel) was plotted versus distance (bottom chart). Note the shift in cytoplasmic intensity profile in FUS^HOMO (gray dots) with respect to FUS^WT (black dots) and FUS^KO (white dots). The histogram on the left represents the mean (+/− SEM) of cytoplasmic intensity between FUS^WT and FUS^HOMO MNs. Scale bar: 10 μm. Results (+/− SEM) are expressed in arbitrary units