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. 2017 Oct 23;7:13749. doi: 10.1038/s41598-017-13664-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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W-2b is cytotoxic and induces premature senescence in cancer cells. (A) Structure of novel isoxazoline derivative of withaferin A (W-2b). (B) Graphs showing the percent cell viability of MCF7, HCT-116 and fR2 cells in response to logarithmic concentrations of W-2b for 24 h, 48 h, and 72 h. (C) Effect of vehicle, doxorubicin (100 nM), WA, and various concentrations of W-2b on SA-β-gal activity in MCF7, HCT-116, and fR2 cells. Original magnification 20x. (D) Bar graph showing quantification of SA-β-gal positive cells. Error bars: mean ± s.d. *P < 0.05. (E) After treatment, MCF7 and HCT-116 cells were stained with DAPI containing mounting media for 15–20 min and observed for the formation of SAHF under Floid Cell Imaging Station. Insets (red boxes) showing magnified images for proper visualization of SAHF. Data are representative of three independent experiments. Original magnification 20x.