5×102 22RV1, C4–2, or 22RV1 Delta-Notch1 cells were plated per well of a six-well plate in triplicate. Cells were treated with DMSO vehicle control, Enzalutamide (5μM), Abiraterone (5μM), DAPT (50μM), or RO4929097 (20μM) (All compounds were purchased from Selleckchem, Houston, TX, catalog numbers S1250, S2246, S2215 and S1575 respectively) (28 (link)–30 (link), 27 (link)). Cells were cultured 9 days, with media and compounds changed every third day. Colonies were then fixed with methanol and stained with 0.1% crystal violet one hour at room temperature. Plates were washed by submerging in a water bath for one hour. Colonies were counted, and colony formation rate determined (percentage), quantified as number of colonies per 5×102 cells X 100, as previously described (21 (link)).
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Prostate Cancer Colony Formation Assay
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Corresponding Organization : Stanford University
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Variable analysis
independent variables
- DMSO vehicle control
- Enzalutamide (5μM)
- Abiraterone (5μM)
- DAPT (50μM)
- RO4929097 (20μM)
- Colony formation rate (percentage), quantified as number of colonies per 5×10^2 cells X 100
- 5×10^2 22RV1, C4–2, or 22RV1 Delta-Notch1 cells plated per well of a six-well plate in triplicate
- Cells were cultured 9 days, with media and compounds changed every third day
- Colonies were fixed with methanol and stained with 0.1% crystal violet
- DMSO vehicle control
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