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. 2020 Oct 17;11(10):874. doi: 10.1038/s41419-020-03085-6

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Differentiated SH-SY5Y cells were treated with amitriptyline (20 μM) or rapamycin (2.5 μM) for 24 h and analyzed LAMP2 and βIII-tubulin localization by immunofluorescence. Scale bar corresponds to 50 μm. Quantification of LAMP2 and DAPI intensity along the arrow line in a is shown in the right panel. b HEK-293T cells were transiently transfected with Arl8-GFP and PLEKHM2 and treated with 20 μM amitriptyline for 24 h. The binding of Arl8 and SKIP/PLEKHM2 in cells was analyzed by immunoprecipitation with anti-GFP and immunoblotting with anti-PLEKHM2. Quantification of PLEKHM2 per Arl8-GFP is shown in the right panel (n = 6). c HEK-293T cells were treated with amitriptyline for 24 h and the binding of KLC1 and PLEKHM2 in cells was analyzed by immunoprecipitation. Quantification of KLC1 per PLEKHM2 is shown in the right panel (n = 3). d Schematic diagram showing the effect of amitriptyline on lysosome positioning. ***P < 0.001. AMI amitriptyline, IP immunoprecipitation, IB Immunoblotting, Rapa rapamycin.