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. 2020 May 1;9(5):1124. doi: 10.3390/cells9051124

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Accumulation of autolysosomes and p62 aggregates by impaired autophagic flux. (a) Autolysosome was determined by the colocalization of the Cyto-ID fluorescence and LysoTracker Red (LTR) in MO3.13 cells treated with/without psychosine (Psy) concomitant with/without chloroquine (CQ) and counter-stained with Hoechst. (b) Quantification of autolysosome puncta in cells. (c) Cytoplasmic deposition of p62 and ubiquitin by chloroquine. Cells were double labeled with anti-p62 and anti-ubiquitin and counterstained with DAPI. Values are expressed as mean ± S.D. (n = 3). Statistical significance *** p< 0.001 in comparison to control (C). C/CQ vs. Psy, or Psy/CQ, ^###p < 0.001. Psy vs. Psy/CQ, ⁺⁺⁺p < 0.001.