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. 2017 Jan 24;3:16050. doi: 10.1038/celldisc.2016.50

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Copyright © 2017 The Author(s)

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Samd4 (sterile alpha motif domain containing protein 4) bound to 5′ untranslated region (UTR) of mitogen-inducible gene 6 (Mig6) and inhibited MIG6 protein translation in C3H10T1/2. For all panels (a–e), top, the effects of Samd4 and Samd4 mutants (K245A R248A K251Q and A281Q) were assessed by analyzing the luciferase activity when luciferase cDNA was fused to the indicated elements. Medium, protein level of Flag-Samd4 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the control measured by western blottings. Bottom, relative expression level of luciferase RNA measured by quantitative reverse transcriptase-PCR. (a–c) Analyses of gradient Samd4 impacted activities of luciferases indicated. Gradient concentrations of SAMD4 were obtained by 0, 50, 200 and 800 ng plasmid transfections, respectively. (d, e) The effects of Samd4 and Samd4 mutants were assessed by analyzing the luciferase activity. EF1α, EF1α promoter; 5′UTR, 5′UTR of Mig6 mRNA. Values represent mean±s.d. (n=3). P-values were obtained from t-tests with paired or unpaired samples. ns, not significant..