Fig. 2.

Inhibition of MEK/ERK and PI3K/Akt signaling pathways down-regulates autophagy. U937 cells were either treated with 10 μM 27-hydroxycholesterol (27-OH) alone or pre-incubated with PD98059 (40 μM) or LY294002 (25 μM) 45 min before 27-OH treatment. Untreated cells were taken as controls. LC3 I and LC3 II, Beclin 1 (A) and p62 (B) protein levels 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 and ##p < 0.01vs. 27-OH.