Figure 2.
2. Anchorage-independent growth and inhibition of anchorage-independent growth by inhibiting MAPK of CREF and various Ha-ras and v-raf transformed CREF cells. (A) Cells were seeded at densities ranging from 1 × 104 to 1 × 105 in 0.4% Noble agar on a 0.8% Noble agar base layer prepared in complete growth medium, fed with 0.4% Noble agar containing medium every 5 to 7 days and colonies >0.1 mm were enumerated after 21 days. (B) Cells were seeded as above in the presence of 0.05% DMSO (control) or 50 µM PD98059, added to the base and overlay layers and added every 5 to 7 days with complete growth medium to the overlay layer. Results represent the averages ± SD for triplicate plates, duplicate assays varied by ≤15%.