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Schematic representation of the Nutlin-3a counter-screen and possible outcomes. Once clones emerge from senescent crisis, the Nutlin-3a counter-screen is performed. Cells are treated for five days with 10 µM Nutlin-3a to determine the response of the immortalised HUF. Mixed and resistant (mutant-TP53) cultures are expanded into immortalised cell lines. Sensitive (WT-TP53) cultures are watched for recovery and discarded if they do not recover within 48 h.
