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. 2020 Nov 5;10:19185. doi: 10.1038/s41598-020-75356-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Olanzapine caused lysosome and autophagosome accumulation in mHypoA-59 neurons in a dose-dependent manner under serum-starved conditions. (A–C) mHypoA-59 neurons were incubated for 24hrs with and without 100 µM olanzapine under serum and serum-starved conditions. After staining with acridine orange (AO), quantification of acidic vesicles were determined as an increase in red (FL3) vs. green (FL1) using flow cytometry (A,B). Flow cytometric assessment of FL3 intensity (C). Respective images of A-B using a live-cell imager. (D,E) mHypoA-59 neurons were incubated for 24 h with or without 100 µM olanzapine under serum and serum-starved conditions. After staining with Cyto-ID, autophagosomes were quantified measuring an increase in green fluorescence (FL1) using flow cytometry (** p < 0.01, ***p < 0.001 compared to the serum only group). (F) mHypoA-59 neurons were incubated for 24hrs with and without 100 µM olanzapine under serum-starved conditions, LC3B-II and p62 were assessed via immunoblotting (* p < 0.05, ***p < 0.001 compared to the serum only group, data is presented as mean ± SEM) Abbreviations: Ola: Olanzapine.