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. Author manuscript; available in PMC: 2013 Dec 9.
Published in final edited form as: Nat Methods. 2007 Aug 12;4(9):10.1038/nmeth1079. doi: 10.1038/nmeth1079

Figure 2.

Figure 2

Efficacy of microRNA sponges. (a–c) RLuc activity relative to firefly luciferase activity was assayed in 293T cells 24 h after transfection with RLuc microRNA target reporters, firefly luciferase transfection control and microRNA sponge plasmids. An RLuc target regulated by 7 miR-20 sites was derepressed by GFP sponges and U6 sponges with bulged or perfect binding sites for miR-20 (a). C, CMV sponge; U, U6 sponge. CX, CXCR4 control; 20b, 7 bulged miR-20 sites; 20pf, two perfect miR-20 sites. Bars represent the expression of the miR-20 target relative to an untargeted control reporter. We measured an artificial CXCR4 target reporter with a single bulged binding site in the presence of a control GFP sponge against miR-21 (miR-21 sponge) or a GFP sponge containing seven CXCR4 binding sites (CXCR4 sponge; b). We transfected cells with 20 nM antisense oligonucleotide (2′ O-methyl 20 or LNA 20) or with the CMV bulged sponge against miR-20 (sponge 20; c). Negative controls; mock (no oligonucleotides or sponges), 2′ O-methyl against miR-30, LNA against miR-122, CXCR4 sponge. We performed each experiment at least three times and have shown a representative example. Error bars, s.d; n = 3.