Featured protocol,
Authors: Sara SdelciSara SdelciAffiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Bio-protocol author page: a4753 
Stefan KubicekAffiliation 1: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, Vienna, Austria
Affiliation 2: Christian Doppler Laboratory for Chemical Epigenetics and Antiinfectives, CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria
For correspondence: skubicek@cemm.oeaw.ac.atBio-protocol author page: a4754
DOI: https://doi.org/10.21769/BioProtoc.2368.
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| Brief version appeared in Nat Chem Biol, Jul 2016 |
The ubiquitously expressed bromodomain-containing protein 4 (BRD4) is an epigenetic reader, which recruits transcriptional regulatory complexes to acetylated chromatin. Because of its role in enhancing proliferation, BRD4 has become a therapeutic target in oncology, as the inhibition of this protein leads to the reduction of the growth of many tumours. Even though BRD4 is more and more studied, its mechanism of action has not been fully described yet. Therefore, we aimed at generating a cellular reporter system to monitor BRD4 inhibition. Such reporter can be potentially used in high throughput chemical and genetic screenings, in order to uncover new possible BRD4 functional pathways. The deeper understanding of the mechanism of action of BRD4 activity will certainly help in developing new therapy strategies for those cancers so called BRD4-dependent.
Featured protocol,
Authors: Feng DuFeng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
Bio-protocol author page: a4714 Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, China
For correspondence: leedyzhao@fmmu.edu.cnBio-protocol author page: a4715 Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
DOI: https://doi.org/10.21769/BioProtoc.2364.
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| Brief version appeared in J Cell Biol, Aug 2015 |
Tumorigenicity refers to the ability of cultured cells to develop viable tumors in immune-deficient animals. The goal of this protocol is to illustrate tumorigenicity assay by subcutaneous tumor-cell-transplantation in nude mice. Target cells are transplanted to 6-week-old nude mice subcutaneously and the tumor growth is monitored over a period of observation or treatment. When tumor grows to a pre-determined size or by the end of the limited period, the nude mice will be euthanatized and the xenograft will removed for further examination.
Featured protocol,
Authors: Thuy L. Phung Thuy L. PhungAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
For correspondence: tphung@bcm.eduBio-protocol author page: a4800 Sriram AyyaswamyAffiliation: Department of Pathology, Texas Children’s Hospital and Baylor College of Medicine, Houston, USA
Bio-protocol author page: a4801
DOI: https://doi.org/10.21769/BioProtoc.2369.
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| Brief version appeared in Cancer Res, Jan 2015 |
To investigate whether endothelial Akt1 activation is sufficient to induce vascular tumor formation in the skin, we have developed a skin graft model in which a skin fragment from transgenic donor mice with inducible and endothelial cell-specific overexpression of activated Akt1 (myrAkt1) is grafted into the skin of wild type recipient mice. The donor skin successfully engrafts after two weeks and, more importantly, vascular tumor develops at the site of transgenic skin graft when myrAkt1 expression is turned on. This skin graft model is a novel approach to investigate the biological impact of a key signal transduction molecule in a temporal, localized and organ-specific manner.
Featured protocol,
Authors: Feng DuFeng DuAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4714 Xiaodi ZhaoAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
For correspondence: leedyzhao@fmmu.edu.cnBio-protocol author page: a4715 Daiming FanAffiliation: State key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China
Bio-protocol author page: a4716
DOI: https://doi.org/10.21769/BioProtoc.2351.
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| Brief version appeared in J Cell Biol, Aug 2015 |
Soft agar colony formation assay is established to estimate the anchorage-independent growth ability of cells. In this assay, a bottom layer of agar with complete media is poured and solidified first, followed by an upper layer containing a specified number of cells suspended in medium-agar mixture. After two weeks of incubation, the number of colonies will be counted, serving as an indicator of malignancy of tumor cells.