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. 2011 Mar 14;108(13):5148–5153. doi: 10.1073/pnas.1102033108

Fig. 3.

Fig. 3.

miR-K5 and miR-K6-3p repress ORF56 protein expression. (A) Genomic DNA expression constructs consist of a 3FLAG tag and the region spanning ORF56 coding region and its entire 3′UTR (solid line between ORF56 and polyadenylation sequence, AATAAA). Defective miRNA binding sites are illustrated as small boxes in the 3′UTR of 3FLAG-56 tK5 and tK6-3p (tK5 and tK6-3p are described in Fig. 2 B and C). ORF57 coding sequence (dashed boxes) and the intron are indicated. (B) Immunoblot analyses of 3FLAG-56 (anti-FLAG immunoprecipitates) and transfection control Firefly luciferase (FLUC) in HEK293 cells either mock transfected (-), or cotransfected with the indicated plasmids and miRNAs for 48 h. Bands labeled NS represent proteins that are nonspecifically precipitated by anti-FLAG antibody. Quantitation of 3FLAG-56/FLUC is described and plotted in Fig. S4A.