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. Author manuscript; available in PMC: 2012 Jul 10.

Published in final edited form as: Oncogene. 2009 Nov 16;29(6):930–936. doi: 10.1038/onc.2009.368

a,b) IMEC cells and c,d) Rat1A fibroblasts expressing the proteins indicated were used in the soft agar transformation assay, performed according to (Cowling et al., 2006) with the following modifications. 2.5 × 10⁴ IMECs/ml were plated in 0.3% noble agar/IMEC medium supplemented with 5% FBS (Invitrogen). Cells were fed with 250μl IMEC medium/5% FBS every 2nd day, starting with the day of plating. Rat1A cells were plated in DMEM/10% FCS and fed every 4th day, starting with the day of plating. a, c) Micrographs of colonies at 14 and 7 days, respectively. b, d) After 7 days the size of colonies was measured for 100 plated cells using a graticle. Chart depicts the percentage of cells developing into colonies of the size indicated at 7 days. Average results for three independent experiments are shown and error bars indicate the standard deviation.

a,b) IMEC cells and c,d) Rat1A fibroblasts expressing the proteins indicated were used in the soft agar transformation assay, performed according to (Cowling et al., 2006) with the following modifications. 2.5 × 10⁴ IMECs/ml were plated in 0.3% noble agar/IMEC medium supplemented with 5% FBS (Invitrogen). Cells were fed with 250μl IMEC medium/5% FBS every 2nd day, starting with the day of plating. Rat1A cells were plated in DMEM/10% FCS and fed every 4th day, starting with the day of plating. a, c) Micrographs of colonies at 14 and 7 days, respectively. b, d) After 7 days the size of colonies was measured for 100 plated cells using a graticle. Chart depicts the percentage of cells developing into colonies of the size indicated at 7 days. Average results for three independent experiments are shown and error bars indicate the standard deviation.