Figure 4.

Interaction of EPG5 with autophagy molecules. (A) LCL of HD, PT1, PT2 and PT3 treated with CpG-FITC and labelled with anti-LAMP2 (red). Colocalization can be seen in yellow pixels (high magnification in insets). Graph shows colocalization between CpG and the lysosomes. Error bars are shown as SEM. (B) EPG5-GFP labeling in healthy fibroblasts did not overlap with EEA1 vesicles distribution, but colocalized with lysosomes stained with LAMP2 antibody (red). (C) 3D surface reconstruction of confocal image stacks of HEK293T cells transfected with a plasmid encoding EPG5-GFP and stained for EPG5 and EEA1 or LAMP2 antibodies showed a significant colocalization between EPG5 and lysosomes (arrows). (D) Z-reconstructions of confocal images (upper panel) of HEK293T cells transfected with a plasmid encoding EPG5-GFP and labeled with anti-LAMP2 antibody. XZ and YZ orthogonal planes (lower panel) of Z-stacks in higher (left) and central (right) focal planes showed that EPG5 clearly colocalized with lysosomes, with their content and/or around them (arrows).