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. 2016 Mar 31;7(3):e2161. doi: 10.1038/cddis.2016.67

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Copyright © 2016 Macmillan Publishers Limited

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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PUMA is the initiator in the Nupr1/Chop axis-activated classical mitochondria apoptotic signaling pathways. HUVECs were transfected with siRNAs targeting Nupr1 (a), Chop (c), PUMA (e) or ctrl siRNA for 48 h followed by METH (1.25 mM) treatment for 24 h. Western blot (a, c, e, j) and quantitative analyses (b, d, f, g–i, k–m) were performed to evaluate the expression of PUMA, Bax, Bcl-2, cleaved-PARP (Cl-PARP), cleaved-caspase-3 (Cl-casp3), the ratio of Bax/Bcl-2, cytosolic cyto c (cyt cyt-c), mitochondrial cyto c (Mit cyt-c) and the Cyt cyt-c/Mit cyt-c ratio. (n) Mitochondrial membrane potential (MMP) was analyzed with fluorescent probe JC-1 assay. (o) Quantitative results of JC-1 assay. Data are presented as mean±S.D. (n=3). *P<0.05 versus the saline vehicle-treated control group and **P<0.05 versus the scrambled+METH group (one-way ANOVA)