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. 2022 Mar 24;12:5076. doi: 10.1038/s41598-022-09078-z

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© The Author(s) 2022

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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AXLi limits autophagic flux at a late stage in the autophagy pathway. Melmet 1 cells were treated with 1 µM AXLi for 48 h with or without co-treatment with 100 nM lysosomal inhibitor Bafilomycin A1 (Baf) for the 3 final hours of the treatment (DMSO for control). As a positive control, cells were in addition to Baf subjected to amino acid starvation by incubation in EBSS medium for 3 h. (a) Expression levels of indicated proteins in the differently treated cells. A representative western image is shown (cropped gel). Quantified expression levels of LC3B-II (after normalization to the loading control, β-actin) presented as relative to non-treated control set to 1. Average ± St. Dev, n = 2. (b) The LDH sequestered assay was performed and the results presented as the mean LDH sequestration activities (in %/h) in differently treated cells, normalized to the control set to 0. Average ± SEM, n = 3, *p ≤ 0.05; **p ≤ 0.01.