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. Author manuscript; available in PMC: 2017 May 1.
Published in final edited form as: Mol Cancer Ther. 2016 Mar 3;15(5):1053–1062. doi: 10.1158/1535-7163.MCT-15-0828

Figure 3. H89 increased ADP-ribosylation of eEF2 and rate of apoptosis.

Figure 3

A. KOPN-8 cells were treated with 10 μM H89 for 1 hour, then, 200 ng/ml of LMB-11 was added for the indicated times before cell lysis, followed by Western Blot analysis for Mcl-1, cleaved PARP, and actin. Not yet modified eEF2 in the cell lysates was labeled in a cell free reaction with biotinylated ADP by LMB-11 as described in methods. B. KOPN-8 cells were treated with 10 μM H89 or vehicle for 1 hour, then left untreated or treated with the indicated concentrations of LMB-11 for 24 (left panel) or 48 hours (right panel), respectively. Cells were then stained with Annexin V-PE and 7-AAD. Living cells were defined as double negative and blotted as percentage of all cells. Standard errors of mean were generated from three independent experiments.