title: iRFP is a real time marker for transformation based assays in high content screening.
abstract: Anchorage independent growth is one of the hallmarks of oncogenic transformation. Here we show that infrared fluorescent protein (iRFP) based assays allow accurate and unbiased determination of colony formation and anchorage independent growth over time. This protocol is particularly compatible with high throughput systems, in contrast to traditional methods which are often labor-intensive, subjective to bias and do not allow further analysis using the same cells. Transformation in a single layer soft agar assay could be documented as early as 2 to 3 days in a 96 well format, which can be easily combined with standard transfection, infection and compound screening setups to allow for high throughput screening to identify therapeutic targets.
authors.affiliation: Cancer Research UK Beatson Institute, Glasgow, United Kingdom.
authors.forename: Viola
authors.initials: V
authors.lastname: Paulus-Hock
authors.affiliation: Cancer Research UK Beatson Institute, Glasgow, United Kingdom.
authors.forename: Eric C
authors.initials: EC
authors.lastname: Cheung
authors.affiliation: Cancer Research UK Beatson Institute, Glasgow, United Kingdom; The Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Patricia
authors.initials: P
authors.lastname: Roxburgh
authors.affiliation: Cancer Research UK Beatson Institute, Glasgow, United Kingdom.
authors.forename: Karen H
authors.initials: KH
authors.lastname: Vousden
authors.affiliation: Cancer Research UK Beatson Institute, Glasgow, United Kingdom.
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Hock
mesh_terms: D016475:3T3 Cells; D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002471:Cell Transformation, Neoplastic; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice
chemical_list: D014408:Biomarkers, Tumor; D008164:Luminescent Proteins