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. 2017 May 17;12(5):e0177901. doi: 10.1371/journal.pone.0177901

Fig 5. Tethering of ZC3H32 to a reporter mRNA represses expression.

Fig 5

(A) Principle of the tethering assay. The reporter mRNA has a chloramphenicol acetyltransferase (CAT) open reading frame. The capped spliced leader is shown as a circle. Five boxB sequences (shown as a single lollipop) bind to the lambdaN peptide. In our case the lambdaN peptide is at the N-terminus of a ZC3H32 open reading frame with a C-terminal myc tag. The negative control reporter (lower diagram) has no boxB. (B) Effect on CAT enzyme expression from tethering lambdaN-ZC3H32-myc to the CAT reporter mRNA. CAT activity is shown as a proportion of that seen in cells with no tethered protein. Results are arithmetic mean ± standard deviation for three clones. (C) Effect on CAT mRNA of tethering lambdaN-ZC3H32-myc in the 3'-UTR. A typical Northern blot is shown. (D) Expression of lambdaN-ZC3H32-myc in the six different clones.