Fig. 4

CLU expression reduces TDP-43 mislocalization in Drosophila neurons. Confocal microscopy images of: a Motor neuron cell bodies of 10-day-old adult Drosophila brains expressing TDP-43 alone (i & ii) or together with CLU (iii and iv). Images (i & ii) and (iii & iv) represent the same fields of view; TDP-43 (red; i & iii), TOTO-3 stained nuclei (grey; ii & iv). In (i), white arrowheads indicate paranuclear foci; in all panels white elipses indicate nuclei (scale bars 5 μm). b Central lobe of adult Drosophila brains expressing TDP-43 +/− CLU. Images in (i) and (iii) were collected using a 40X objective and encompass the entire central lobe (scale bars 50 μm); elipses show motor neuron cell bodies and the central region of each image contains neuronal process and axons. Images in (ii) and (iv) represent optical zooms of the areas indicated by the white boxes in (i) and (iii), respectively (scale bars 5 μm). TDP-43 (red), white arrowheads in (ii) indicate axons. c Motor neuron cell bodies of adult Drosophila brains co-expressing CLU (green) and TDP-43 (red); TOTO-3 (white; nucleoli stain intensely, the rest of the nucleus stains less brightly); (iv) overlay of all three images, yellow indicates co-localisation of CLU and TDP-43 (scale bars 7 μm). d Motor neuron cell bodies of third instar larval Drosophila brains co-expressing CLU (green) and TDP-43 (red), TOTO-3 (blue); (iv) overlay of all three images; yellow indicates co-localisation of CLU and TDP-43 (scale bars 5 μm). e Reducing Western blot of the soluble and insoluble fractions of 10-day-old adult Drosophila head homogenate comparing non-transgenic (non-TG) Drosophila and Drosophila expressing TDP-43 +/− CLU. β-actin was used as a loading control. Results shown are representative of several independent experiments