Figure 9.

Analysis of parental and PTEN-deleted (clones 16 and 22) Hec1A cells by immunoblot analysis (A). Deletion of PTEN activated AKT in clones 16 and 22. Both clones 16 and 22 expressed reduced cyclin D1 levels under normal growth conditions. Growth of parental cells was affected by high (10%) or low (1%) serum, whereas serum had no effect on growth of clone 16 (C). All time points for the proliferation assay were done in triplicate, and the values represent results from three independent experiments. Immunoblot analysis of PTEN, p-AKT, p-ERK, and cyclin D1 in whole-cell lysates of serum-starved cells after adding serum for indicated time periods (B) and represent the results obtained from three independent experiments. Addition of 10% serum increased cyclin D1 levels in the parental cells by at least threefold to fourfold as early as 30 minutes (D).