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. 2006 Dec 27;1(1):e116. doi: 10.1371/journal.pone.0000116

Figure 4. Luciferase activity assay.

Figure 4

Luciferase reporter constructs were generated through placing fragments of 185 bp (Construct I, sequence from nt31–216 of the VEGF 3′-UTR) and 241 bp (Construct II, sequence from nt703–944 of the VEGF 3′-UTR) at the 3′ end of luciferase gene in pRL-TK. COS-7 cells were co-transfected with a luciferase reporter vector and an miRNA which has a putative binding site in either Construct I (A) or Construct II (B). Luciferase activity was measured to determine the effects of these miRNAs on luciferase translation. In addition to a random sequence (NC), miR-29, miR-150, and miR-383 which have putative binding sites in the 3′-UTR of VEGF but not on Construct I or II, as predicted by all of the algorithms, were employed as negative controls. *, p<0.05; **, p<0.01.