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. 2016 Mar;22(3):373–382. doi: 10.1261/rna.054833.115

FIGURE 4.

FIGURE 4.

TTP mutated in tetraprolines 1 and 2 is impaired in its ability to repress a luciferase reporter ARE-mRNA. (A) Luciferase luminescence assays from TTP−/− MEFs transiently coexpressing wild-type or mutant TTP with two reporters, one encoding firefly luciferase (F-Luc) containing in its 3′ UTR the AU-rich element (ARE) from human GM-CSF mRNA, and one encoding Renilla luciferase (R-Luc) without an ARE as an internal control. For each sample, F-Luc activity was normalized to R-Luc and values were normalized to the wild-type TTP-transfected condition, which was set as 1. Error bars represent standard error of the mean (SEM) (n = 4). P-values were calculated using Student's t-test (paired, two-tailed). (B) Representative Western blot of samples used in panel A. PABP was used as the loading control. (C) Luciferase assays in 4EHP+/+ and 4EHP−/− MEFs similar to the experiment in panel A except in the absence of cotransfected TTP and the absence or presence of transiently expressed 4EHP and GYF2 as indicated. Error bars represent SEM (n = 3).