Figure 5
Nrf2 activators restore the p62 deficient phenotype. (A) NADH redox index was evaluated after incubation with the Nrf2 activators TBE-31(TBE, 20 nM), sulphoraphane (SFN, 10 μM) and the synthetic triterpenoid RTA-408 (RTA, 50 nM). Cells were plated on 25 mm coverslips in 6 well plates. When they reached 70% confluency, cells were treated 24 hours with TBE, SFN and RTA separately. NADH redox index was calculated as the basal level relative to maximal respiration after FCCP (1 mM) (0%) and inhibited respiration after NaCN (1 mM) (100%) (see Fig. 2B) in fibroblasts from patients carrying the A381V and K238del pathogenic mutations in p62 compared to two control fibroblasts (C1, C2) in presence of the Nrf2 activators. All data represents the mean of at least 3 independent experiments ±SEM. In all cases ** indicates p < 0.01 and *** indicates p < 0.001 compared with the values in control cells. (B) NADH pool expressed as absolute values between maximal and minimal respiration (see Fig. 2B) in control and patient fibroblasts in presence of the Nrf2 activators. Experimental conditions were the same as in (A). All data represents the mean of at least 3 independent experiments ±SEM. In all cases *** indicates p < 0.001 compared with the values in control cells. (C) The ΔΨm was analyzed in fibroblasts from patients and controls upon activation of Nrf2. Experimental conditions were the same as in (A). All data represents the mean of at least 3 independent experiments ±SEM. In all cases ** indicates p < 0.01 and *** indicates p < 0.001 compared with the values in control cells.
