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. Author manuscript; available in PMC: 2011 Jun 1.
Published in final edited form as: Int J Biochem Cell Biol. 2010 Mar 20;42(6):1030–1038. doi: 10.1016/j.biocel.2010.03.012

Figure 4. Phosphorylation of p21 increases its stability.

Figure 4

Figure 4

A) Phospho mimics of p21 increase steady levels of p21 in HCT116 cells. HCT116 cells were transfected with equal amounts of one of the HA-tagged p21 plasmids (WT, T/D, S/D, T/A, S/A, DD or AA). Forty hours post transfection, cells were harvested and the steady level of the p21 proteins were evaluated by Western blot with anti-HA antibody. The membrane was reprobed with anti-actin antibody to verify equal loading. B) Stability assay of various p21 proteins. HCT116 cells were transfected with one of the HA-tagged p21 plasmids (DD, T/D, S/D, WT, S/A, T/A, or AA). Thirty-six hours after transfection, cells were treated with 25μg/ml cycloheximide (CHX) and harvested at indicated time points. The stabilities of p21 and actin proteins were analyzed by Western blot with anti-p21 and anti-actin antibodies.