FISH detection of miR-31* expression in OLK and mtOLK.

<p>(<b>A</b>) ∼ (<b>C</b>), no positive signal is detected in OLK. (<b>D</b>) ∼ (<b>L</b>), the representative positive signals are showed in mtOLK. (<b>D</b>) ∼ (<b>F</b>), tumor nest; (<b>G</b>) ∼ (<b>I</b>), vascular area; (<b>J</b>) ∼ (<b>L</b>), inflammation. FISH signals are visualized in green, while blue depicts nuclear DAPI stain. OLK, oral leukoplakia; mtOLK, malignant transformed oral leukoplakia.</p

Dual luciferase reporter assay for miR-31* and targeted genes.

<p>(<b>A</b>) Psicheck-FGF3-wt/mt and miR-31*; (<b>B</b>) Psicheck-IL5Rα-wt/mt and miR-31*. NC  =  negative control; mimic  =  miR-31* mimic; wt  =  wild type; mt  =  mutant type.</p

Representative results of miR-31* effects on biological functions.

<p>(<b>A</b>) the MTT assay in HIOEC; (<b>B</b>) the Annexin V assay in Leuk-1; (<b>C</b>) flow cytometry analysis for cell cycle in Cal-27; (<b>D</b>) representative field of view of HIOEC invasion inserts at a 100× magnification, (Da) miR-31* mimic, (Db) miR-31* inhibitor, (Dc) negative control, (Dd) Quantification of relative numbers of invading cells representing average counts from 6 fields-of-view per insert per sample ±SD, **p<0.05, ***p<0.001; (<b>E</b>) representative field of view of HIOEC migration inserts at a 100× magnification, (Ea) miR-31* mimic, (Eb) miR-31* inhibitor, (Ec) negative control, (Ed) quantification of relative numbers of invading cells representing average counts from 6 fields-of-view per insert per sample ±SD, **p<0.05, ***p<0.001.</p

Clinical outcomes in OLK and mtOLK.

<p>OLK, oral leukoplakia; mtOLK, malignant transformed oral leukoplakia; ND, no dysplasia; MiD, mild dysplasia; MoD, moderate dysplasia; SD, severe dysplasia; SCC, squamous cell carcinoma; NA, not available.</p

miR-31* mimics/inhibitors regulating mRNA and protein expression of FGF3 in oral cells.

<p>(<b>A</b>) QRT-PCR analysis of FGF3 expression in Leuk-1, HIOEC and Cal-27 cell lines, **p<0.05; (<b>B</b>) Western blot analysis of of FGF3 expression in Leuk-1, HIOEC and Cal-27 cell lines transfected with miR-31* mimic, miR-31* inhibitor and negative control. The representative blot is shown (upper for FGF3). The histogram shows the average volume density normalized by the loading control, GAPDH (lower).</p

Unsupervised hierarchical cluster analysis of miRNA expression in OLK and mtOLK.

<p>The marked separation of OLK and mtOLK based on the miRNAs profiling is exhibited. Scale is shown in the upper right. Red represents a high expression and green represents a low expression. OLK, oral leukoplakia; mtOLK, malignant transformed oral leukoplakia.</p