Figure 3
From: t-DARPP regulates phosphatidylinositol-3-kinase-dependent cell growth in breast cancer
t-DARPP expression enhances anchorage-independent cell growth in soft agar. A, MCF-7 cells stably expressing t-DARPP or pcDNA3.1 empty vector were seeded in soft agar onto 6-well tissue culture plates (105 cells per well). The cells were grown for one month to allow for anchorage-independent colony formation. B, Quantification of colony formation indicated that MCF-7/t-DARPP cells produced markedly higher number of colonies than control cells (>4-fold increase, p < 0.001). C, Protein extracts from MCF-7/t-DARPP and MCF-7/pcDNA3.1 cells were analyzed by western blot for t-DARPP, p-AKT (S473), AKT, p-GSK3β (S9), GSK3β, C-Myc, Cyclin D1, and β-actin. t-DARPP expression directly correlated with increased phospho-AKTser473, phospho-GSK3βser9, C-Myc, and Cyclin D1 protein levels. Results are representative of at least three experiments and shown as the mean ± SD. Significance of difference was calculated using Student's t test analysis.