Figure 5.
Neurons exposed to HNE exhibit accumulations of polyubiquitinated proteins. (A) Representative images showing ubiquitinated protein immunoreactivity in neurons exposed to vehicle (Con) or 5 µM HNE for 6 hours. Scale bar, 10 µm. (B) Puncta of ubiquitinated protein aggregates were quantified in 22 neurons in control cultures and 30 neurons in HNE-treated neurons in 3 separate experiments. ***p<0.001. (C, E, I J and K) Total protein from neurons treated with ethanol or HNE for 6 hours (C, E and G) or 24 hours (I and K) were separated in 4–12% Bis-Tris gels, and the proteins were then transferred onto a nitrocellulose membrane. Representative examples of blots stained with Ponceau S (left) and immunoreactivity with the indicated antibodies (right) are shown. (D, F, H, J and L) Results of densitometric analyses of immunoblots from 3 separate experiments. **p<0.01, ***p<0.001.