Figure 4

Specificity of the anti-FUS IgG and co-localization of hFUS with NPM-hMLF1 in eye imaginal disc cells. (a) Western immunoblotting analysis. Proteins were extracted from adult heads carrying GMR-GAL4 > UAS-GFP (GMR-GAL4/Y; UAS-GFP/+) and GMR-GAL4 > UAS-hFUS (GMR-GAL4/Y; UAS-hFUS/+). The blots were probed with anti-hFUS IgG. The bands with an apparent molecular mass of 83 kDa and 72 kDa correspond to hFUS protein. α-tubulin was used as a loading control. The flies were developed at 28 °C. Three independent experiments were carried out. (b–i and b’–i’) Immunostaining of eye imaginal discs with the anti-hFUS IgG. (b–e and b’–e’) GMR-GAL4 > UAS-lacZ (GMR-GAL4/Y; UAS-lacZ/+). (f–i and f’–i’) GMR-GAL4 > UAS-hFUS (GMR-GAL4/Y; UAS-hFUS/ + ). Eye imaginal discs were stained with DAPI (b,f,b’ and f’), phalloidin (c,g,c’ and g’) and anti-hFUS IgG (d,h,d’ and h’). Merged confocal images are shown in (e,i,e’ and i’). Higher magnification images of the indicated regions in the panels (b–i) are shown in (b’–i’) respectively. Arrowheads indicate MF. a indicates anterior and p indicates posterior. The flies were developed at 28 °C. Bars indicate 50 μm (b-i) and 5 μm (b’–i’). (j–r and j’–q’) Immunostaining of eye imaginal discs with the anti-hFUS IgG and the anti-hMLF1 IgG. (j–m and j’–m’) GMR-GAL4 > UAS-hFUS/UAS-lacZ (GMR-GAL4/Y; UAS-hFUS/UAS-lacZ+) (n–r and n’–q’) GMR-GAL4 > UAS-hFUS/UAS-NPM-hMLF1 (GMR-GAL4/Y; UAS-hFUS/UAS-NPM-hMLF1). Eye imaginal discs were stained with DAPI (j,n,j’ and n’), anti-hFUS IgG (k,o,k’ and o’) and anti-hMLF1 IgG (l,p,l’ and p’). Merged confocal images are shown in m,q,r,m’ and q’. Higher magnification images of the indicated regions in the panels (j–q) are shown in (j’–q’) respectively. Arrowheads indicate MF. a indicates anterior and p indicates posterior. The flies were developed at 28 °C. Bars indicate 50 μm (j–q) and 5 μm (r and j’–q’).