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. 2017 May 4;7:1498. doi: 10.1038/s41598-017-01659-7

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© The Author(s) 2017

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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Treatment with cryptolepine inhibits mitochondrial biogenesis and associated molecular targets in melanoma cells. (a) Mitochondrial biogenesis in cryptolepine-treated and non-treated A375 and Hs294t cells was determined by measuring the levels of SDH-A and COX-I protein expression using the Mitochondrial Biogenesis Kit according to the manufacturer’s protocol. The experiment was performed two times. Protein expression in the vehicle-treated control group was considered as 100%. Statistical significance versus control, ^¶ P < 0.01 and ^† P < 0.001. (b) After treatment of cells with cryptolepine for 24 h, total cell lysates from A375 and Hs294t cells were prepared and subjected to western blot analysis to determine the levels of proteins involved in mitochondrial biogenesis. Equal loading of proteins was verified by stripping the membrane and reprobing with β-actin antibody. (c) Immunofluorescence staining for the detection of Opa1-positive and SIRT1-positive cells was performed as detailed in Materials and Methods. Photomicrographs were obtained using Keyence Fluorescence Microscope BZ-X710 (Keyence Corporation of America). Representative photomicrographs are shown.