Posttranscriptional Gene Regulation by Spatial Rearrangement of the 3′ Untranslated Region
Figure 4
Suppression of EJC-Enhanced NMD by Poly(A) Tail FB
(A) Schematic illustration of the mRNAs expressed by the indicated constructs. The 26-nucleotide sequence located 28 nucleotides downstream of codon 310 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. Predicted EJCs in the 3′ UTR are shown by yellow ovals. WT = construct with full-length ORF; ter310 = construct with PTC at codon 310; SL = stemloop control construct with complementary sequence inserted in exon C2.
(B) Half-lives of the FB mRNAs were measured as described in Figure 1C.
(C) Relative miniμ mRNA levels from FB constructs with 3′ UTR introns expressed in cells with (+) or without (−) Upf1 knockdown were measured as in Figure 2C and are shown below the histogram. The histogram depicts the fold increase of miniμ mRNA upon Upf1 knockdown.
(D) The Upf1 knockdown efficacy was assessed by Western blotting as in Figure 2D. Detection of Sm B/B' served as loading control.