Figure 4
p62 deficiency results in increased oxidative stress. (A,B) Cellular oxidative stress was evaluated analysing the oxidation rates of the superoxide indicator dihydroethidium (Het) which initially exhibits cytosolic blue fluorescence and when oxidised, it shows bright red fluorescence in the nucleus. These changes can be detected by live-cell imaging. Time-course representative traces showing the Het oxidation slopes in untransfected, SCR and p62 KD SH-SY5Y cells (A) and fibroblasts from control donors and patients carrying the p62 pathogenic mutations mentioned above (B). In all cases the traces represent the mean of at least 20 cells on a single coverslip ±SEM. (C,D) Het oxidation rates from untransfected, SCR and p62 KD SH-SY5Y cells (C) and control and p62 mutant fibroblasts (D). Rates were calculated as the change in fluorescence over a period of time and are expressed as percentage of a basic rate. Data represent the mean of at least 3 independent experiments ±SEM. In all cases * indicates p < 0.05 compared with the values in control cells.
