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. 2015 Sep 25;9:5323–5335. doi: 10.2147/DDDT.S90859

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© 2015 Rathore and Cekanova. This work is published by Dove Medical Press Limited, and licensed under Creative Commons Attribution – Non Commercial (unported, v3.0) License

The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.

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AD198 (AD)- and DOX-induced apoptosis and caspase activation in tested K9TCC and K9OSA cell lines in vitro.

Notes: (A) K9TCC#2-Dakota and K9OSA#1-Zoe cells were treated with 1 μM DOX and 1 μM AD for 24 hours and induced apoptosis was measured using the TACS Anexin V-FITC assay using a flow cytometer. Relative apoptotic activities were normalized to control groups. Values represent mean ± standard error (n=9) of three replicates of three independent experiments. Paired Student’s t-tests comparing treatment to control groups (*P<0.05, **P<0.01, ***P<0.001) and comparing among DOX and AD treatment groups (^##P<0.01, ^###P<0.001) were used. (B) K9TCC#2-Dakota and K9OSA#1-Zoe cells were treated with 1 μM DOX and 1 μM AD for 24 hours and caspase activities were measured using the Caspase-Glo 3/7 luminescence assay. Relative caspase activities were normalized to control. Values represent mean ± standard error (n=6) of two replicates of three independent experiments. Paired Student’s t-tests comparing treatment to control (***P<0.001) and comparing among DOX and AD treatments (^##P<0.01) were used. (C) K9TCC#2-Dakota and K9OSA#1-Zoe cells were treated with 1 μM DOX and 1 μM AD for 24 hours. The expressions of PARP(CF) were evaluated by WB analysis. Actin was used as loading control. The results are representative of three independent experiments (n=3). (D) Densitometry evaluation of PARP protein bands from WB analysis was done using ImageJ software. Values represent the mean of measured densitometry of each protein’s band from two replicates of three independent experiments ± standard error (n=6). Paired Student’s t-tests were used to compare controls to DOX and AD treatments (*P<0.05, ***P<0.001) and to compare DOX to AD treatment (^##P<0.01).

Abbreviations: AD, AD198; PARP(CF), PARP-cleaved fragment; DOX, doxorubicin; K9OSA, canine osteosarcoma; K9TCC, canine transitional cell carcinoma; PARP, poly (ADP-ribose polymerase); PARP(T), total PARP; WB, Western blot.