Evaluating Anticancer Effects of BITC

To perform clonogenicity assay42 (link), breast cancer cells were treated with BITC as indicated for 10-days; colonies were counted. Anchorage-independent growth of breast cancer cells in the presence of BITC was assayed by colony formation in soft agar43 (link). Cell viability assay was performed using a commercially available XTT assay kit (Roche Applied Science, Indianapolis, IN). Mammosphere assays were performed as previously described44 (link) and spheres (>50 μm) were counted. For apoptosis analysis, cells were stained with Annexin-V and propidium iodide (PI) followed by fluorescence-activated cell sorting (FACS) analysis.

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Xie B., Nagalingam A., Kuppusamy P., Muniraj N., Langford P., Győrffy B., Saxena N.K, & Sharma D. (2017). Benzyl Isothiocyanate potentiates p53 signaling and antitumor effects against breast cancer through activation of p53-LKB1 and p73-LKB1 axes. Scientific Reports, 7, 40070.

Publication 2017

XTT assay kit

Annexin v Apoptosis Assays Cancer of breast Cell viability Cells Propidium iodide

Corresponding Organization :

Other organizations : Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, University of Maryland, Baltimore, Semmelweis University, HUN-REN Research Centre for Natural Sciences

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Variable analysis

independent variables

BITC treatment

dependent variables

Clonogenicity
Anchorage-independent growth
Cell viability
Mammosphere formation
Apoptosis

control variables

Not explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

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