Figure 3

TM-Grb2, TM-Shc1 and TM-Shc2 oncoproteins induce anchorage-independent growth and anoikis resistance in IEC-6 cells. (A) The ability of the indicated IEC-6 cell populations to grow in an anchorage-independent manner was tested in soft agar assays. The bar graph represents the average number of colonies formed in soft agar, expressed as percentage ± S.E.M. of those produced by Tpr-Met-IEC-6 cells, from 3 independent experiments performed in triplicate. Tpr-Met-IEC-6 cells, when seeded at a density of 5000 cells in a 6-well plate, were producing an average of 300–500 colonies after ~ 10 days. (B) Photographs depict typical morphology of the colonies formed in soft agar. (C) The bar graph represents the average size of colonies formed in soft agar, expressed as percentage ± S.E.M. of those produced by Tpr-Met-IEC-6 cells, from 3 independent experiments performed in triplicate. (D) Anoikis sensitivity of these IEC-6 cell populations was tested. Cell viability was measured by XTT assays 18 hours after seeding the cells in suspension or adherent conditions. The bar graph shows in percentage the mean ± S.E.M. value of viable cells grown in suspension relative to the adherent ones, calculated from 4 independent experiments performed in triplicate. (E) Cell viability in adherent condition, expressed as the percentage mean ± S.E.M. relative to Tpr-Met, is shown.