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Figure 3 | BMC Molecular Biology

Figure 3

From: Cell type-dependent gene regulation by Staufen2 in conjunction with Upf1

Figure 3

Tethering of Stau2 to the 3'-UTR of a reporter mRNA enhances its protein expression in 293F cells. (A) Schematic representations of the reporter constructs. The reporter Rluc-8xMS2bs consists of a cDNA encoding the Renilla luciferase (Rluc) protein with eight repeats of the MS2 binding site (8xMS2bs) in its 3'-UTR. The control plasmid Fluc contains a cDNA encoding the firefly luciferase (Fluc) protein. The expression of both mRNAs is under the control of the herpes simplex virus thymidine kinase (TK) promoter. (B-E) MS2-fusion proteins indicated below each panel were co-expressed in HeLa (B, C) or 293F cells (D, E) with Rluc-8xMS2bs and Fluc. At 48 h after transfection, the cells were harvested and subjected to Western blotting (B, D) or luciferase assay (C, E). (B, D) Western blotting was performed using an anti-HA antibody to detect MS2-fused proteins. (C, E) The expression of Rluc relative to that of Fluc was calculated for each transfection, and the normalized level of the MS2 transfection was defined as 1. Values are expressed as the means ± standard error of the mean (SEM) of four independent experiments.

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