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. 2018 Nov 27;9:5006. doi: 10.1038/s41467-018-07344-1

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© The Author(s) 2018

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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Fig. 2 — HSV-P10 AKT inhibition. a Western blots of lysates from the indicated cells infected with HSVQ (Q) or HSV-P10 (P) or left untreated (NV). Infected cells (MOI = 1) were lysed 24 hpi and probed for expression of PTENα, pAKT (S473), Akt, and GAPDH. L indicates the moleculcar weight ladder. b Western blot for secreted PTEN expression in conditioned medium. Culture media was concentrated 20X from U87∆EGFR cells infected 24 h with either HSVQ (Q) or HSV-P10 (P10) and probed for secreted PTENα. L indicates the molecular weight ladder. c Flow cytometric analysis of pAkt-S473 and virus encoded GFP in infected U87∆EGFR cells 15 hpi. d Spectral analysis of fluorescent intensity along a random line drawn across the view field of one or two cells visualizing pAKT (red), DAPI (blue) and GFP (green) of the indicated cells. y-axis represents arbitrary fluorescence units (AFU). Note the higher intensity of pAKT in HSVQ-infected (GFP positive) cells relative to adjacent uninfected (GFP negative) cell (middle panel). Note the lack of increased pAKT intensity in HSV-P10 infected (GFP positive) cells relative to adjacent uninfected (GFP negative) cells (bottom panel)