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. 2012 Sep 28;31(21):4236–4246. doi: 10.1038/emboj.2012.271

Figure 1.

Figure 1

ZAP inhibits the translation of target mRNA. (A) Schematic representation of Nef-luc mRNA. (B) 293TREx-ZAP cells were infected with VSV-G pseudotyped NL4-3-luc virus for 3 h, mock-treated (0) or treated with tetracycline at a final concentration of 20 or 1000 μg/ml for additional 48 h to induce ZAP expression. One-tenth of the cells were lysed and luciferase activity was measured (open bars). The luciferase activity in mock-treated cells was set as 100. Cytoplasmic RNA was extracted from the remaining cells and levels of Nef-luc mRNA were measured by real-time PCR (black bars). The levels of Nef-luc mRNA normalized to that of GAPDH mRNA from mock-treated cells were set as 100. (C) Translation efficiency of Nef-luc mRNA was calculated as luciferase activity divided by Nef-luc mRNA level in (B). Translation efficiency of Nef-luc mRNA in mock-treated cells was set as 1. (D) Schematic representation of Luc-Mk mRNA. (E) 293TREx-ZAP cells were transfected with the pCMV-FL-Mk reporter and then mock-treated (0) or treated with tetracycline at a final concentration of 20 or 1000 μg/ml for 48 h to induce ZAP expression. One-tenth of the cells were lysed and luciferase activity was measured (open bars). The luciferase activity in mock-treated cells was set as 100. Cytoplasmic RNA was extracted from the remaining cells and levels of Luc-Mk mRNA were measured by real-time PCR (black bars). The levels of Luc-Mk mRNA normalized to that of GAPDH mRNA from mock-treated cells were set as 100. (F) Translation efficiency of Luc-Mk was calculated as luciferase activity divided by Luc-Mk mRNA level in (E). Translation efficiency of Luc-Mk in mock-treated cells was set as 1. Data presented are mean values±s.d. from three independent experiments.