Lack of LC3–GFP clustering in neurons expressing mutant CHMP2B.

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Lack of LC3–GFP clustering in neurons expressing mutant CHMP2B. Lack of LC3–GFP clustering in neurons expressing mutant CHMP2B. Neurons were co-transfected at 10 DIV with the indicated CHMP2B plasmids (left), together with the autophagosomal marker LC3–GFP. The cells were fixed 5 days later and processed for immunofluorescence staining with anti-tag antibody and Alexa-Fluor-594-coupled secondary antibody. Confocal images were acquired in both the immunofluorescence and GFP channels. Diffuse LC3–GFP localisation predominated upon co-transfection with CHMP2Bwt (a1,a2), CHMP2BΔ10 (c1,c2) and in a large majority of cases with CHMP2Bintron5 (b1,b2). Autophagosome accumulation was seen in rare cells transfected with CHMP2Bintron5 (b3, b4, arrow). Arrowhead: no autophagosomes were detected in the dendrites. Scale bars: 10 μm. Agnès Belly et al. J Cell Sci 2010;123:2943-2954 © 2010.