Fig. 5.

Autophagosomes produced both in proband fibroblasts and in healthy control group (father and brother) (a). Double immunocytochemical staining of fibroblast cells showed colocalization of LC3 with p62. LC3 protein was visualized in red, p62 in green, nuclei in blue, and autophagosomes in yellow. A yellow signal signifies colocalization of LC3A and p62. In order to better visualize the fluorescent signals present within the large merged images, zoom images were shown. In proband fibroblast increased the number of autophagy markers; LC3A (b) and p62 (c) were analyzed using ImageJ. Production of autophagosomes was shown healthy fibroblast cells and proband fibroblast cells. Colocalization analysis of LC3 with p62-labeled lysosomes was performed on ImageJ based on Coloc 2 analysis and Pearson value (d). The data are represented as the mean ± S.E.M. One-way ANOVA was used for statistical analysis (*p < 0.05, **p < 0.025)