Figure 1.

Autophagy degradation phase impairment in Familial Alzheimer’s disease associated to presenilin 1 A246E mutation (FAD1) fibroblasts. (A) Representative Western blot of Microtubule-associated protein 1 light chain 3 (LC3) expression for the study of autophagy flux as explained in “Materials and Methods” Section and Supplementary Figure S1, in control and FAD1 fibroblasts treated or not with carbonyl cyanide m-chlorophenylhydrazone (CCCP; 20 μM) in the absence or presence of NH₄Cl (15 mM). (B,C) Quantification of LC3II levels (B) and LC3II/LC3I ratio (C) in FAD1 cells with respect to the control ones under basal conditions. (D,E) Quantification of LC3II synthesis (D) and degradation (E) ratios as described in “Materials and Methods” Section. (F,G) Western blot of p62 expression after the treatment as in (A) and quantification of basal levels (G). (H,I) Quantification of p62 accumulation (H) and degradation (I) ratios (n = 3 independent experiments using the control/Alzheimer’s disease (AD) fibroblast couple AG04148/AG06840; ^†p < 0.08; *p < 0.05; **p < 0.01).