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. 2015 Feb 17;9:993–1026. doi: 10.2147/DDDT.S73493

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© 2015 Wang et al. 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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CDDO-Me-induced apoptotic death in Ec109 and KYSE70 cells determined by measuring cellular DNA contents using flow cytometry.

Notes: (A) Percentages of specific cell populations showed in dot plots and apoptotic cells showed in bar graphs for Ec109 and KYSE70 cells treated with CDDO-Me at 0.25–1.0 μM for 24 hours. (B) Percentages of specific cell populations showed in dot plots and apoptotic cells showed in bar graphs for Ec109 and KYSE70 cells treated with CDDO-Me at 0.5 μM for 4–48 hours. (C) Representative blots of respective proteins measured to show the effect of CDDO-Me treatment on the expression levels of Bcl-xl, Bcl-2, Bax, PUMA, cytochrome c, cleaved caspase-3 (active), cleaved caspase-9 (active), PARP, and cleaved PARP in Ec109 and KYSE70 cells determined using Western blotting analysis. (D) Bar graphs showing the levels of Bcl-xl, Bcl-2, Bax, PUMA, cytochrome c, cleaved caspase-3 (active), cleaved caspase-9 (active), PARP, and cleaved PARP in Ec109 and KYSE70 cells. Data represent the mean ± SD of three independent experiments. β-actin was used as the internal control. *P<0.05, **P<0.01, and ***P<0.001 by one-way ANOVA and Tukey’s post hoc test to compare the levels of the protein measured with the control cells treated with control vehicle only (0.05% DMSO, v/v).

Abbreviations: CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; SD, standard deviation; ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; PARP, poly ADP ribose polymerase; PUMA, p53 upregulated modulator of apoptosis; 7-AAD, 7-amino-actinomycin D; PE, phycoerythrin; hr, hour.

CDDO-Me-induced apoptotic death in Ec109 and KYSE70 cells determined by measuring cellular DNA contents using flow cytometry.

Notes: (A) Percentages of specific cell populations showed in dot plots and apoptotic cells showed in bar graphs for Ec109 and KYSE70 cells treated with CDDO-Me at 0.25–1.0 μM for 24 hours. (B) Percentages of specific cell populations showed in dot plots and apoptotic cells showed in bar graphs for Ec109 and KYSE70 cells treated with CDDO-Me at 0.5 μM for 4–48 hours. (C) Representative blots of respective proteins measured to show the effect of CDDO-Me treatment on the expression levels of Bcl-xl, Bcl-2, Bax, PUMA, cytochrome c, cleaved caspase-3 (active), cleaved caspase-9 (active), PARP, and cleaved PARP in Ec109 and KYSE70 cells determined using Western blotting analysis. (D) Bar graphs showing the levels of Bcl-xl, Bcl-2, Bax, PUMA, cytochrome c, cleaved caspase-3 (active), cleaved caspase-9 (active), PARP, and cleaved PARP in Ec109 and KYSE70 cells. Data represent the mean ± SD of three independent experiments. β-actin was used as the internal control. *P<0.05, **P<0.01, and ***P<0.001 by one-way ANOVA and Tukey’s post hoc test to compare the levels of the protein measured with the control cells treated with control vehicle only (0.05% DMSO, v/v).

Abbreviations: CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; SD, standard deviation; ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; PARP, poly ADP ribose polymerase; PUMA, p53 upregulated modulator of apoptosis; 7-AAD, 7-amino-actinomycin D; PE, phycoerythrin; hr, hour.