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. 2017 Nov 23;7:16159. doi: 10.1038/s41598-017-16327-z

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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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Glutaminase protein expression in PC3 and PC3M cell lines before and after CB-839 treatment. PC3 and PC3M cells were treated (+) or not treated (−) with GLS1 inhibitor CB-839 (1 μM) and their expression of GLS1 and GLS2 proteins were determined by Western blot. Antibodies specific for GAC and KCA were utilized on two replicate samples, while GLS2 expression was determined on three replicates. PC3 cell lysates had higher levels of expression of GLS2 than PC3M cell lysates regardless of CB-839 treatment. The average density of bands in the GLS2 blots were determined by using ImageJ software. The statistical significance was determined by unpaired two-tailed t-test. The difference in GLS2 expression was significant in the untreated cells (P < 0.03) but because of high variability the difference was not significant in the drug-treated cells. Levels of GLS1 (GAC and KGA) were similar in all cell lysates. The full gels can be seen in the supplemental data.