Figure 4

miR-196a specifically binds to the 3' UTR of bovine NOBOX and regulates its expression. (A) Schematic of the luciferase reporter constructs used to demonstrate sequence specificity in recognition sequence in bovine NOBOX 3' UTR mediating miR-196a mediated repression. Nucleotides changed to generate the target site mutant 3' UTR are underlined (B) Repression of luciferase activity due to specific interaction between miR-196a and the predicted MRE in the luciferase-NOBOX-3' UTR constructs. Repression of luciferase reporter gene activity by miR-196a was abolished when the MRE was mutated. Data is presented as relative firefly luciferase units (RLUs). Relative firefly luciferase values were determined by a ratio of firefly to renilla luciferase with the negative control (cells transfected with native NOBOX-3' UTR construct alone) set at 1. Each group represents the mean ± SEM of four wells for an experiment repeated four times with similar results. Different letters indicate statistical difference (P < 0.05).