Cells were seeded at a density of 1–2 × 103 cells/well in a 6-well plate and treated with increasing concentrations of X15695 and cultured for 14–21 days. Medium and compounds were exchanged after 7 days. Cells were fixed with methanol/acetic acid mixture [3:1 volume for volume (v/v)] and the formation of colonies was visualized using 0.5% crystal violet (w/v) in 20% methanol (v/v). Plates were scanned in a conventional office scanner (Epson). The area covered by colonies was calculated using the ColonyArea Plugin for ImageJ (RRID:SCR_003070; ref. 24 (link)).
Pan M., Solozobova V., Kuznik N.C., Jung N., Gräßle S., Gourain V., Heneka Y.M., Cramer von Clausbruch C.A., Fuhr O., Munuganti R.S., Maddalo D., Blattner C., Neeb A., Sharp A., Cato L., Weiss C., Jeselsohn R.M., Orian-Rousseau V., Bräse S, & Cato A.C. (2023). Identification of an Imidazopyridine-based Compound as an Oral Selective Estrogen Receptor Degrader for Breast Cancer Therapy. Cancer Research Communications, 3(7), 1378-1396.
Other organizations :
Karlsruhe Institute of Technology, Nantes Université, Inserm, University of British Columbia, Institute of Cancer Research, Royal Marsden NHS Foundation Trust, Dana-Farber Cancer Institute, Harvard University
Cell fixation (methanol/acetic acid mixture [3:1 volume for volume (v/v)])
Colony visualization (0.5% crystal violet (w/v) in 20% methanol (v/v))
Image acquisition (conventional office scanner)
controls
No positive or negative controls were explicitly mentioned.
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