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. 2019 Sep 25;9:13902. doi: 10.1038/s41598-019-50157-5

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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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Etoposide-mediated apoptosis is mitigated in GRK2 overexpressing cells. (A–D) Tva-expressing UW228 (A,B) and Daoy cells (C,D) were infected with pRDAV-mycGRK2-IRES-eGFP virus to express human GRK2 or the control virus pRDAV-eGFP, grown for 8 days and then treated with etoposide (10 µg/ml for UW228, 8 µg/ml for Daoy) for the indicated times. Flow cytometry was used to assess apoptosis (AnnexinV/7AAD) in the eGFP-expressing cells. (B,D) are means ± SEM of early + late apoptosis of three biological replicates for each of these two cell lines; p-values: in B * represents p = 0.04 and ** represents p = 0.007; in D * represents p = 0.03 and ** represents p = 0.001, using unpaired student’s t-test. (E,F) Western blot for PARP1 cleavage in lysates of UW228 (E) or Daoy cells (F) infected with GRK2 or control vectors as in (A–D) that were treated with etoposide (10 µg/ml for UW228; 8 µg/ml for Daoy) for the indicated times. Supplemental Figure S4 provides a GRK2 western blot of cells used in (E,F) with longer exposure, to show native GRK2 level compared to overexpressed.