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. 2017 Jan 10;7:40070. doi: 10.1038/srep40070

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Copyright © 2017, The Author(s)

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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(A) Total protein lysates of LKB1-depleted (LKB1^shRNA) and vector control (pLKO.1) MCF7 cells were immunoblotted for the expression of LKB1. MCF7-LKB1^shRNA and MCF7- pLKO.1 cells were treated with 2.5 μM BITC and total protein lysates were examined for LKB1 expression in an immunoblot assay. β-actin was used as control. (B) MCF7-LKB1^shRNA and MCF7- pLKO.1 cells were treated with 2.5 μM BITC and subjected to XTT assay. *p < 0.01, compared with untreated controls. Vehicle-treated cells are denoted with the letter “C”. (C) Total protein lysates from MCF7-LKB1^shRNA and MCF7- pLKO.1 cells treated with 2.5 μM BITC for 8 and 16 hours were immunoblotted for the expression of cleaved PARP and total PARP. β-actin was used as a control. Bar diagram shows quantitation of western blot signals from multiple independent experiments. (D) MCF7-LKB1^shRNA and MCF7- pLKO.1 cells were treated with 2.5 μM BITC and subjected to Annexin V/PI staining. *p < 0.01, compared with untreated controls. (E) MCF7-LKB1^shRNA and MCF7- pLKO.1 cells were treated with 2.5 μM BITC and subjected to soft-agar colony formation assay. *p < 0.01, compared with untreated controls. (F) MDA-MB-231-LKB1^shRNA and MDA-MB-231-pLKO.1 cells were treated with 2.5 μM BITC and subjected to soft-agar colony formation assay. *p < 0.01, compared with untreated controls. (G) LKB1-depleted (LKB1^shRNA) and vector control (pLKO.1) MDA-MB-231 cells derived tumors were developed in nude mice and treated with vehicle and BITC. Tumor growth was monitored by measuring the tumor volume for 6 weeks. (n = 8–10); (p < 0.001), pLKO.1+ BITC compared with LKB1^shRNA + BITC. (H) Tumors from LKB1^shRNA+Vehicle, pLKO.1+Vehicle, LKB1^shRNA + BITC, and pLKO.1+ BITC groups were subjected to Immunohistochemical (IHC) analysis using p21, PUMA and DR5 antibodies.