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. 2017 Feb 2;8(2):e2594. doi: 10.1038/cddis.2017.6

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Copyright © 2017 The Author(s)

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(a) Intracellular levels of ROS were determined in A-375 at 1, 2, 4 and 24 h of treatment with 30 μM vemurafenib (Vem), 50 μM TRAM-34 (TRAM) and the combination. The assay is based on staining with the ROS-sensitive dye H₂DCFDA and subsequent flow cytometry. (b) Examples of treated cells versus controls are given. H₂O₂-treated cells served as positive control; cells with high ROS are indicated. The experiment was performed in triplicates; at 2 and 4 h, at least two independent experiments revealed highly comparable results. (c) Increased ROS levels are shown in Mel-2a at 2 h in response to vemurafenib (30 μM), TRAM-34 and the combination. Examples are shown on the right side (triplicate determinations, two experiments)