Effects of binding site mutagenesis on miR-142-3p-induced repression of <i>Bmal1</i> 3′ UTR activity.

<p>(A) Diagrammatic representation of WT and mutagenized <i>Bmal1</i> 3′ UTR constructs with independent site-directed deletions of predicted miR-142-3p binding sites. (B) Normalized bioluminescence from HEK293 cells co-transfected with pEZX-MR04 miR-142 expression vector and with either the target control, full-length <i>Bmal1</i> (WT), <i>Bmal1 c.1_7del, Bmal1 c.335_357del</i> or <i>Bmal1 c.1_7del</i>+<i>c.335_357del</i> miRNA 3′ UTR target clones. (C) Normalized bioluminescence from HEK293 cells co-transfected with pEZX-MR04 miR-494 expression vector with either the target control, full-length <i>Bmal1</i> (WT), <i>Bmal1 c.1_7del, Bmal1 c.335_357del</i> or <i>Bmal1 c.1_7del</i>+<i>c.335_357del</i> miRNA 3′ UTR target clones. Bars represent mean (±SEM) determinations of luciferase bioluminescence for each treatment group (n = 4). The plotted values correspond to the ratios of firefly luciferase signal normalized to <i>Renilla</i> luciferase activity in the same sample and are represented as a percentage of the average signal for control vector transfectants. Asterisks denote comparisons in which normalized bioluminescence for a given treatment group was significantly reduced (<i>p</i><0.05) relative to that observed in control vector transfectants.