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. 2013 Dec 19;289(6):3164–3175. doi: 10.1074/jbc.M113.524413

FIGURE 5.

FIGURE 5.

Rbm24 stabilizes p21 transcript. A–C, ectopic expression of Rbm24 has no obvious effect on the level of p21 pre-mRNA. MCF7 (A), HCT116 (B), and HCT116 (p53−/−) (C) cells were uninduced (−) or induced (+) with 0.5 μg/ml tetracycline to express Rbm24 for 48 h. Total RNAs were isolated and then subjected to RT-PCR analysis to examine the level of p21 pre-mRNA. GAPDH pre-mRNA was measured as a control. D and E, the level of p21 pre-mRNA remains relatively even upon knockdown of Rbm24. H1299 (D) and HCT116 (p53−/−) (E) cells were transfected with a lentivirus containing either control luciferase shRNA (shluc) or Rbm24 shRNA (shRbm24) and then selected by puromycin (1 μg/ml) for 3 days prior to RT-PCR analysis to determine the level of p21 pre-mRNA. GAPDH pre-mRNA was measured as a control. F, HCT116 (p53−/−) cells were treated with actinomycin-D (5 μg/ml) over an 8-h period at 2-h intervals. Total RNAs were purified, and the levels of p21 and actin transcripts were analyzed using RT-PCR. G, the relative levels of p21 transcript were normalized with the levels of actin transcript and plotted along with time to calculate the relative half-life of p21 mRNA in the presence or absence of Rbm24. The x axis represents the time after addition of actinomycin-D (5 μg/ml), and the y axis represents the relative levels of remaining p21 transcript in the cells.