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. 2007 Jun 4;27(15):5365–5380. doi: 10.1128/MCB.00113-07

FIG. 7.

FIG. 7.

Defining the exact nucleotide sequence of the HuR-BS within the region 2A. (A) Scheme of the nucleotide sequence of the 2A region within the 3′UTR of β-actin. The three new probes, 2AL, 2AM, and 2AR, that cover the 2A region are shown. (B) HuR associates with the 2AR region but not with the 2AL or the 2AM region. Gel shift binding assay performed by incubating 500 ng of purified GST or GST-HuR protein with radiolabeled 2AL, 2AM, and 2AR. The HuR-complex is indicated (HuR-C). (C) The unlabeled 2AR but not the 2AM element is able to compete away the association of HuR with the full-length β-actin 3′UTR. Gel shift binding assay performed by incubating purified protein GST or GST-HuR with radiolabeled full-length 3′UTR, as well as a 1/10×, 1×, 10×, or 100× excess of unlabeled 2AR (specific) or 2AM (nonspecific). The asterisk shows the HuR-RNA complex.