Figure 4.

Determination of AURKA 3′UTR stability in neuroblastoma cells overexpressing MCPIP1 protein. Relative AURKA transcript levels following the treatment with (A) actinomycin D and (B) DMSO. For the determination of statistical significance two-way analysis of variance with Tukey’s post-hoc test was performed. Relative luciferase activity of BE(2)-C (C) and KELLY (D) cells co-transfected with MCPIP1-wt, MCPIP1-D141N or control expression vectors and pmiRGlo-AURKA_3′UTR reporter vector calculated compared to the relative activity of corresponding cells transfected with an empty pmiRGlo vector. Relative luciferase transcript levels in BE(2)-C (E) and KELLY (F) cells transfected with MCPIP1-wt, MCPIP1-D141N or control expression vectors and pmiRGlo-AURKA_3′UTR reporter vector assessed by RT-qPCR. Renilla luciferase was used as a reference. All experiments were performed three times. Graphs represent mean ± SEM. The one-way analysis of variance was implemented with Tukey’s post hoc test. C, cells co-transfected with the empty expression vector and pmiRGlo-AURKA_3′UTR reporter vector; MCPIP1-wt, cells transfected with wild type MCPIP1 expression vector and pmiRGlo-AURKA_3′UTR reporter vector; MCPIP1-D141N, cells transfected with mutated MCPIP1 expression vector and pmiRGlo-AURKA_3′UTR reporter vector.*P < 0.05, **P < 0.01; ^ P < 0.05, ^^^ P < 0.001; && P < 0.01