Fig. 4.

Modulation of autophagy induction by N-acetyl cysteine (NAC) and diphenyleneiodonium chloride (DPI). Effect of NAC or DPI on 27-OH-dependent p62 induction: cells were pre-incubated (1 h) with 100 μM NAC (A) or 50 μM DPI (B) and then treated with 27-OH (10 μM) for 6 h. Effect of NAC on 27-OH-dependent autophagy induction: cells were pre-incubated (1 h) with 100 μM NAC and then treated with 10 μM 27-OH for 72 h (C). Untreated cells were taken as controls. Protein levels of p62, LC3 I and LC3 II, and Beclin 1 were analyzed by Western blotting. One blot representative of three experiments is shown. Histograms represent the mean values ± S.D. of three independent experiments; LC3 I, LC3 II, Beclin 1, and p62 band intensities were quantified using ImageJ software; densitometric measurements were normalized against the corresponding actin levels and expressed as percentage of control value *p < 0.05 and **p < 0.01 vs. control group; #p < 0.05 vs. 27-OH.