Fig. 6.

Translation attenuates miR-K5-mediated down-regulation. (A) Experimental constructs for dual luciferase assays. 3′UTRg (with an intron, used in Figs. 1 and 2) and 3′UTR (without an intron, used in Fig. 5) are described previously. In N-ORF, Exon 1 and part of Exon 2 were in-frame fused to Rluc that allows translating an ORF57-Rluc chimeric fusion protein. N-ORF-tK5 is an isogenic derivative of N-ORF in which the miR-K5 site is mutationally inactived, as illustrated by a red symbol at the mutated target site. (B) HEK293 cells were cotransfected with the indicated reporter constructs and miRNAs. Mean values and error bars (SD) were derived from quadruplicate independent transfections. Statistical significance is indicated (p value, two-tailed t test).