Figure 4From: The cyclin-like protein Spy1/RINGO promotes mammary transformation and is elevated in human breast cancer Spy1 mediated colony formation is Cdk1 dependent. 293 cells transfected with the indicated constructs were (A) immunoprecipitated for HA tagged-Cdk1 and blotted for Myc and HA tag. n = 3 (B) imaged after 14 days in a soft agar assay. Transfections and protein levels were monitored by western blot analysis (inset). n = 3 (C) Quantification of colony number over three separate transfections. (D) Soft agar assays of cells transfected in triplicate with Myc-Spy1-TST in the presence or absence of Cdk dominant negative constructs (Cdk1-DN or Cdk2-DN). n = 3 Plates are imaged and quantified at day 14. Errors bars reflect SD between triplicate experiments. ANOVA with post hoc analysis using Tukey test was performed;* P ≤ 0.05, ** P ≤ 0.01.Back to article page