Figure 3
Altered FAD homeostasis is reflected in p62 deficient cells. (A) Time-course representative traces of FAD autofluorescence from untransfected, SCR and p62 KD SH-SY5Y cells. Addition of FCCP (1 μM) maximised respiration and thereby increased FAD autofluorescence to maximal levels (100%). Addition of NaCN (1 mM) then inhibited respiration and reduced the FAD autofluorescence to a minimum (0%). The traces represent the mean of at least 20 cells on a single coverslip ±SEM. (B) Graphical description of the FAD homeostasis analysis by monitoring the FAD autofluorescence using confocal microscopy in p62 deficient cells compared to controls. FAD redox indexes were obtained by calculating the initial FAD autofluorescence when the minimum FAD autofluorescence is normalised to 0% and the maximum to 100%. The FAD redox index generation (the initial redox level expressed as percentage of the range) and the FAD pool are described graphically. (C) FAD redox indexes from untransfected, SCR and p62 KD SH-SY5Y cells representing the mean of at least 3 independent experiments ±SEM. In all cases * indicates p < 0.05 compared with the values in the corresponding control cells. (D) The FAD pool was expressed as absolute values between maximal and minimal respiration in untransfected, SCR and p62 KD SH-SY5Y cells. Data represent the mean of at least 3 independent experiments ±SEM. * indicates p < 0.05 compared with the values in the corresponding control cells.
