Fig. 2. Effects of Euro 5 and Euro 4 DEPs, before any after-treatment, at 3.3 μg/cm² on autophagy: evaluation of LC3-II and p62 levels as autophagic markers in Normal Human Bronchial Epithelial (NHBE) cells.

a Western blot analysis of LC3-II and p62 levels in lysates from NHBE cells (i) untreated and cultured for 48 h with (ii) lysosomal protease inhibitors E64d and Pepstatin A (PepA), (iii) Euro 5, (iv) Euro 5 + E64d + PepA, (v) Euro 4, (vi) Euro 4 + E64d + PepA. Blot shown is representative of nine experiments (N = 9). b Densitometry analysis of LC3-II levels relative to β-actin (mean ± SD). P = 0.0008 for Euro 5 vs untreated; P = 0.0101 for Euro 5 vs Euro 5 + E64d + PepA; P = 0.0022 for Euro 4 vs untreated. c Densitometry analysis of p62 levels relative to β-actin (mean ± SD). No significant (NS, P > 0.05) differences in p62 levels were found