Figure 2.
HNE increases LC3 and p62 protein levels in neurons. (A – C) Cortical neurons were treated with 0.05% ethanol (Con) or 5 µM HNE with or without 100 µM bafilomycin A1 for the indicated time periods. Immunoblots of neuronal cell lysates were analyzed using antibodies against LC3I/II and p62, and β–actin (loading control). Representative blots are shown in panel A, and results of densitometric analysis of the LC3II, p62 and β–actin bands are shown in panels B and C. Values (mean and SEM) are from 4 independent experiments. (D – F) Total cellular lysates from neurons subjected to the indicated individual or combined-treatments (5 µM HNE, 100 nM rapamycin and 100 nM bafilomycin A1) were subjected to immunoblot analysis using antibodies against LC3, p62 and β-actin. Representative blots are shown in panel D, and results of densitometric analysis of the LC3II, p62 and β–actin bands are shown in panels E and F. Values (mean and SEM) are from 4 independent experiments. *p<0.05, **p<0.01, ***p<0.001 compared to the control value.