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. 2013 May 10;288(24):17832–17843. doi: 10.1074/jbc.M112.448977

FIGURE 5.

FIGURE 5.

Non-stop mRNA decay requires Dis3. A, down-regulation of Dis3 expression. HeLa cells were transfected with either control luciferase siRNA (lane 1) or Dis3 siRNA-1 (Dis3(#1); lane 2). Total cell lysate was analyzed by Western blotting (WB) using the indicated antibodies. B, down-regulation of Dis3 expression inhibits fast degradation of non-stop mRNA. HeLa cells were transfected with p5×FLAG-EGFP, pT7-TR, either pFBG control (lanes 1–5 and 11–15) or pFBG non-stop (lanes 6–10 and 16–20), and either control luciferase siRNA (lanes 1–10) or Dis3 siRNA-1 (lanes 11–20). The transcriptional pulse-chase analysis was performed as described in the legend to Fig. 2. C, the levels of FBG mRNAs were quantitated as described in the legend to Fig. 2. D, the half-lives of FBG mRNAs were calculated. The asterisks denote the levels of statistical significance (Student's t test) between luciferase and Dis3 siRNAs: *, p < 0.05;**, p < 0.01.