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. 2020 Oct 28;10:18515. doi: 10.1038/s41598-020-75598-1

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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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Reduction of autophagic flux in Sal-treated SK-Mel-19 cells. (a) Samples from DMSO-treated control and Sal-treated (1.0 μM) cells immunoblotted for P62 (*p < 0.05; **p < 0.01; n = 3). (b) Immunofluorescent assay showed the expression of P62 in SK-Mel-19 cells treated with or without 1.0 μM Sal for 24 h. (c) The expression level of P62 was examined by immunohistochemical analysis in 10 mg/kg Sal-treated SK-Mel-19 tumor xenograft. (d) Samples treated with 1.0 μM Sal for the indicated period of time or MG132 (5 μM) for 6 h or CQ (6.25 μM) for 6 h immunoblotted for ubiquitinated proteins. (e) SK-Mel-19 cells was treated with Sal alone, inhibitor alone, or their combination for 24 h. Level of protein expression of LC3B was measured by western blot analysis (**p < 0.01; n = 3).