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. 2017 Aug 24;8(8):e3016. doi: 10.1038/cddis.2017.384

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Copyright © 2017 The Author(s)

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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Compound induced cell apoptosis. (a) SMMC-7721 cells treated with the title compound (20, 40, and 80 nm) for 48 h were stained with Annexin V/PI and then were analyzed using fluorescence cytometry and (b) fluorescence microscopy (magnification × 200). (c) Mitochondrial membrane depolarization analysis was performed by detecting the mitochondrial transmembrane potential using JC-1 stain treated with the compound for 48 h and then it was analyzed with fluorescence cytometry and (d) fluorescence microscopy (magnification × 200). (e) Western blot and densitometric analyses for Bcl-2 /Bax and cyt-c after SMMC-7721 cells treated with the title compound. (f) Compound induced nuclear chromatin condensation. SMMC-7721 cells were treated with 0, 20, 40, and 80 nm of the title compound for 48 h and then were stained with DAPI as described in Materials and Methods. Cells were examined and photographed using a fluorescence microscope at × 200. Representative blots were from three independent experiments. For the statistics of each panel in this figure, data are expressed as mean±S.D. (n=3); scale bars are 50 μm; *P<0.05 versus control, **P<0.01 versus control, ***P <0.001 versus control

Compound induced cell apoptosis. (a) SMMC-7721 cells treated with the title compound (20, 40, and 80 nm) for 48 h were stained with Annexin V/PI and then were analyzed using fluorescence cytometry and (b) fluorescence microscopy (magnification × 200). (c) Mitochondrial membrane depolarization analysis was performed by detecting the mitochondrial transmembrane potential using JC-1 stain treated with the compound for 48 h and then it was analyzed with fluorescence cytometry and (d) fluorescence microscopy (magnification × 200). (e) Western blot and densitometric analyses for Bcl-2 /Bax and cyt-c after SMMC-7721 cells treated with the title compound. (f) Compound induced nuclear chromatin condensation. SMMC-7721 cells were treated with 0, 20, 40, and 80 nm of the title compound for 48 h and then were stained with DAPI as described in Materials and Methods. Cells were examined and photographed using a fluorescence microscope at × 200. Representative blots were from three independent experiments. For the statistics of each panel in this figure, data are expressed as mean±S.D. (n=3); scale bars are 50 μm; *P<0.05 versus control, **P<0.01 versus control, ***P <0.001 versus control