Fig. 4. Serum and anchorage dependence assays of IKLF-transfected cells.
A, cells from the two IKLF·HA-transfected clones (I3 and I10) were seeded at a density of 2 × 105 cells/well in 6-well plates and maintained in 1% FBS for up to 6 days after seeding. Cells were counted every other day as described in the legend to Fig. 3 and plotted over time. The two empty vector-transfected clones (V3 and V5) failed to grow (not shown). B, Ha-ras-, IKLF·HA-, and empty vector-transfected cells were seeded in agar suspension, which were fed DMEM with 10% FBS every other day for 3 weeks. Colonies of cells were scored by visual inspection using an inverted phase-contrast microscope. Shown are the mean colony numbers in 8 random fields under a × 100 magnification. Bars indicate S.E. *, p < 0.005 compared with empty vector-transfected cells (V3 and V5). C, photomicrograph of a typical colony of Ha-ras- and IKLF·HA-transfected cells (left versus middle, respectively). Empty vector-transfected cells (right) produced no such colonies.