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Suppression of EJC-Independent NMD by Poly(A) Tail FB

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posted on 29.04.2008, 00:58 by Andrea B Eberle, Lukas Stalder, Hansruedi Mathys, Rodolfo Zamudio Orozco, Oliver Mühlemann

(A) Schematic illustration of the mRNAs expressed by the indicated constructs. The 26-nucleotide sequence located 42 nucleotides downstream of codon 440 is depicted in red, and the insertion upstream of the poly(A) tail of this sequence (red) or of the complementary sequence (green) is indicated. WT = construct with full-length ORF; ter440 = construct with PTC at codon 440.

(B) Half-lives of the FB mRNAs were measured as described in Figure 1C.

(C) Relative miniμ mRNA levels from the EJC-independent FB constructs shown in (A) normalized to β-globin WT mRNA from a cotransfected plasmid, were measured by RT-qPCR from Upf1-depleted cells (light gray bars) or from control cells expressing a scrambled shRNA (dark gray bars).

(D) The efficacy of the Upf1 knockdown was assessed by Western blotting. Detection of lamin A/C served as loading control.


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