Figure 6

Tax increases the half-life of p21 mRNA. A, HeLa/18 × 21-EGFP cells were transduced with LV-puro, LV-Tax, LV-M22, and LV-M47 as described in METHODS. (Left panels) EGFP expression of cells transduced with the respective vectors. (Right panels) Levels of expression of various Tax alleles in transduced cells as determined by immunoblotting with the 4C5 Tax hybridoma antibody (Tax) and a control antibody against β-actin (actin). B, Tax stabilizes p21 mRNA. HeLa/18 × 21-EGFP cells were transduced with LV-Tax, or LV-puro for 48 h as in (A), and then incubated with actinomycin (2 μg/ml) for the indicated durations. The level of p21 mRNA in each sample was measured by real-time RT-PCR, quantified using the amount of the 18S rRNA as an internal reference, and finally, normalized to the p21 transcript level at time 0 and plotted and analyzed as in Fig. 5. C, APC/C-deficiency correlates with attenuation in p21 mRNA stabilization. (Left panel) HeLa/18 × 21-EGFP cells were transduced with LV-M47, LV-M22, and LV-L235F as in (A). The rate of p21 mRNA turn-over in the transduced cells was determined as in (B). Data analysis and graphing were as described in Fig. 5 A (Right panels). Expression of the respective Tax alleles is as shown in the immunoblot.