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. Author manuscript; available in PMC: 2018 Mar 1.
Published in final edited form as: J Neurochem. 2017 Feb 16;140(6):941–954. doi: 10.1111/jnc.13957

Figure 3.

Figure 3

HNE causes the accumulation of LC3- and p62-immunoreactive puncta in neurons without affecting LC3 and p62 mRNA levels. (A) Neurons exposed to ethanol (Con) or HNE for 6 hours were fixed with 100% cold methanol for 15 minutes at −20°C, and then co-stained with anti-LC3 (green) and anti-p62 (red) antibodies. Nuclear DNA was stained with Hoechst (blue). Images were acquired using a 40× objective with Zeiss LSM 410 confocal microscope. Representative neurons (arrows) from each group are shown in high magnification images on the right. Scale bars: 50 µm in left panels and 10 µm in right panels. (B and C) Numbers of LC3- and p62-positive puncta per neuron were counted. Results were from analyses of 182 control neurons and 62 HNE-treated neurons in three independent experiments (n = 3). ***p<0.001. (D and E) Levels of LC3 (D) and p62 (E) mRNAs were quantified using real-time PCR in RNA samples from neurons treated with ethanol or HNE for the indicated time periods.

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