AKT activity in S and G2 phases: (A) NIH3T3 cells were synchronized with thymidine treatment in S phase and released by thymidine removal for the indicated times. In addition, quiescent cells were left unstimulated (G0), or stimulated with serum for the indicated times. In each case, lysates were collected and probed with an antibody specific for the activating phosphorylation on AKT. This was compared to western analysis of total AKT protein. Cells enter G2 phase 4–5 hrs following thymidine removal. (C) The levels were quantitated, and the ratio of phospho-specific AKT divided by total AKT levels is presented. (B) For comparison, the levels of cyclin D1 were analyzed by western analysis in the same lysates, (D) and quantitated.
Yang et al. BMC Cell Biology 2006 7:33 doi:10.1186/1471-2121-7-33