Figure 5.

c-MycS causes anchorage-independent growth. (a) Anchorage-independent growth of Rat1a cells induced by c-Myc–ER proteins. The stable Rat 1a cell lines expressing c-Myc2–ER, c-MycS–ER, c-MycΔ106–143–ER (ΔMyc–ER) or pMV7 vector alone were plated at 1 × 10⁴ cells/35-mm dish in soft agar containing DMEM plus 10% FCS with or without 2 μm OHT. Colonies (>100 μm) were counted by an Omnicon colony counter (Bausch & Lomb) on the tenth day after plating. (b) Anchorage-independent growth of Rat 1a cells caused by constitutive c-Myc proteins. Rat 1a cells or the stable Rat 1a cell lines expressing c-Myc2, c-MycS, c-MycΔ150, or the pBabepuro vector alone were plated at 1 × 10⁴ cells/35-mm dish in soft agar containing DMEM plus 10% FCS. Colonies (>100 μm) were counted as described above.