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. 2015 Mar 30;16(4):7112–7132. doi: 10.3390/ijms16047112

Figure 1.

Figure 1

Figure 1

HuR binds U-rich regions of the cyclin E1 3'UTR. (A) MCF7 cells were transfected with pcDNA3.1 (vec) or pcDNA3.1 myc-HuR (HuR) or (B), with control siRNA (si-ctrl) or HuR siRNA (si-HuR). 48–72 h after transfection, protein was extracted for western blotting for Cyclin E1 or HuR (top). Values in the graphs are the mean fold change ± SD from three independent experiments. ** p < 0.01 versus control; (C) Sequence of cyclin E1 3'UTR with two HuR binding sites (underlined with light grey) and two miR-16 target sites (underlined with dark grey), as well as a schematic depiction of the cyclin E1 mRNA including the 5'UTR, coding region (CR) and 3'UTR. FL (full length cyclin E1 3'UTR) and (A–E) are 3'UTR segments used for UV cross-link and UV cross-link competition experiments shown in panels D and E; (D) The binding of GST-HuR with the cyclin E1 3'UTR detected by UV cross-link analysis; and (E) Specific or non-specific binding of GST-HuR with the cyclin E1 3'UTR measured by UV cross-link competition using E1CR378 (bases 184–378 in the coding region) as non-specific competitor. Representative crosslinks of at least three replicates are shown in panels D and E.