miR-181a-directed repression of Renilla luciferase reporter genes containing RalA-3′-UTR segments.

<p>K562 cells were cotransfected with scrambled RNA or miR-181a together with RalA-3′-UTR or RalA-mut-3′-UTR luciferase reporter in the presence of firefly luciferase reporter plasmid as indicated. Renilla luciferase activity and firefly luciferase activity were measured by dual-luciferase reporter assay (Promega). Renilla luciferase activity was normalized to firefly luciferase activity. The data represent the mean value of three independent experiments. (A) overexpression of miR-181a significantly repressed Renilla luciferase activity in cells transfected with RalA-3′-UTR, but not RalA-mut-3′-UTR vector; *p<0.01. (B) Human RalA-3′-UTR and its miR-181a target site predicted by TargetScan software. Mut: contains 7-base-mutation at the miR-181a target region.</p

miR-181a regulates cell cycles partially by targeting RalA.

<p>K562 cells were transfected with scrambled RNA, miR-181a, or RalA siRNA and harvested 48 h post-transfection. The cells were stained with PI solution and analyzed using flow cytometry. Both miR-181a and RalA siRNA significantly induced G2-phase arrest. *<i>p</i><0.01, when compared with blank or SCR controls.</p

Downregulated genes upon miR-181a overexpression.

<p>miR-181a targets were identified by combining TargetScan software prediction and expression profiling. The four genes identified by both methods were considered as candidate targets of miR-181a in K562 cells.</p

miR-181a promotes apoptosis partially by targeting RalA.

<p>K562 cells were transfected with scrambled RNA, miR-181a, or RalA siRNA and harvested 48 h after transfection. The cells were stained with FITC-conjugated annexin V and PI, followed by flow cytometry analysis. Annexin V-positive and PI negative cells represent apoptotic cells. *<i>p</i><0.01, when compared with blank or SCR controls.</p

KEGG pathway analysis of RalA downstream regulated genes.

<p>The list of known potential targets of RalA graphed using cytoscape.</p

miR-181a inhibits K562 cells growth partially by targeting RalA.

<p>K562 cells were transfected with scrambled RNA, miR-181a, or RalA siRNA. Viability of the cells was assessed by MTT (<b>A</b>) and trypan blue exclusion assay (<b>B</b>) in triplicates. Both miR-181a and siRNA RalA effectively inhibited cell viability in a time-dependent manner. *<i>p</i><0.01, when compared with blank or Scramble controls.</p

miR-181a regulates RalA expression at the post-transcriptional level.

<p>K562 cells were transfected with scrambled RNA, miR-181a, or RalA siRNA and harvested 48 h after transfection. Total RNA was isolated and analyzed for the expression of miR-181a (A), RalA mRNA (B) and RalA protein (C) using qPCR and Western blot, respectively. *<i>p</i><0.01, when compared with controls.</p

Newcastle-Ottawa quality assessment scale^*.

<p>Newcastle-Ottawa quality assessment scale^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0165179#t001fn001" target="_blank">*</a>}.</p

Quality assessment of included studies.

<p>Quality assessment of included studies.</p

Characteristics of the articles included in the meta-analysis.

<p>Characteristics of the articles included in the meta-analysis.</p