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. 2019 Apr 12;9:6005. doi: 10.1038/s41598-019-42228-4

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© The Author(s) 2019

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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GB elicited phosphorylation of both Akt and MAPK/ERK kinases in pre-B Nalm-6 cells in a time-dependent manner. Whole cell lysates were subjected to Western blot analysis with the antibodies indicated in material and methods. (a,b) The bar graphs represent densitometry analyses showing a comparable increase in pAkt (Ser 473) and pMAPK/ERK phosphorylation induced after 1 h and 3 h of GB-treated cells (50 µl), respectively (n = 2). As a reference control, lysates from untreated (Unt) pre-B Nalm-6 cells were included, and also, to establish the basal levels of Akt and MAPK/ERK phosphorylation; representative Western blots displaying GB-stimulated pAkt (Ser 473), pAkt (Thr 308) and pERK1/2 phosphorylation are shown in the lower panels. (c) The bar graph represents densitometry analysis showing a comparable increase in STAT5 (Tyr 694) phosphorylation caused after 1 h and 3 h of GB-treated cells (50 µl), respectively (n = 3). As a reference control, lysates from untreated (Unt) pre-B Nalm-6 cells were included; western blots are displaying GB-stimulated STAT5 phosphorylation, as compared with total STAT5 protein are shown in (c), lower panel. The 50 µl of GB correspond to 1.5 ± 0.048 mg/ml of GB lyophilized powder.