The constructs of GUS expression driven by <i>AhLEC1B</i> promoter and schematic representation of the different length promoters with 5′ or 3′ terminal deletion.

<p>Q1-Q6 indicates the different promoters with 5′ or 3′ terminal deletion. The white and gray rectangles show the upstream promoter region from TSS and 5′ UTR region respectively.</p

Effects of <i>AhLEC1B</i> promoter deletions on the expression profile of <i>GUS</i> gene in transgenic Arabidopsis lines.

<p>Q1-Q6 indicate the GUS expression patterns in different transgenic Arabidopsis lines containing 5′ or 3′ terminal deletion promoters, and the CK-N and CK-P showed the GUS expression profiles in non-transformed negative control and in positive control harboring 35S:GUS constructs, respectively.</p

Cloning and Characterization of 5′ Flanking Regulatory Sequences of <i>AhLEC1B</i> Gene from <i>Arachis Hypogaea</i> L.

<div><p>LEAFY COTYLEDON1 (LEC1) is a B subunit of Nuclear Factor Y (NF-YB) transcription factor that mainly accumulates during embryo development. We cloned the 5′ flanking regulatory sequence of <i>AhLEC1B</i> gene, a homolog of <i>Arabidopsis LEC1</i>, and analyzed its regulatory elements using online software. To identify the crucial regulatory region, we generated a series of GUS expression frameworks driven by different length promoters with 5′ terminal and/or 3′ terminal deletion. We further characterized the GUS expression patterns in the transgenic <i>Arabidopsis</i> lines. Our results show that both the 65bp proximal promoter region and the 52bp 5′ UTR of <i>AhLEC1B</i> contain the key motifs required for the essential promoting activity. Moreover, <i>AhLEC1B</i> is preferentially expressed in the embryo and is co-regulated by binding of its upstream genes with both positive and negative corresponding <i>cis</i>-regulatory elements.</p></div

The primers used in this study.

<p>The primers used in this study.</p

MiR-377 is downregulated in HCC cell lines.

<p>(A) Downregulation of miR-377 expression was in HCC cell lines. Relative expression of miR-377 in four HCC cell lines (HepG2, SMMC7721 Hep3B and Bel7402) and one normal human hepatocyte (HL-7702) was determined by qRT-PCR. U6 snRNA was used as internal control. (B) Northern blot analysis of miR-377 in four HCC cell lines (HepG2, SMMC7721 Hep3B and Bel7402) and one normal human hepatocyte (HL-7702). U6 snRNA was also detected as a loading control.</p

Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the <i>Arabidopsis</i> NF-YB family.

<p>Phylogenetic tree for peanut AhLEC1A and AhLEC1B, and the <i>Arabidopsis</i> NF-YB family.</p

The sequence of 5′ flanking regulation region of peanut <i>AhLEC1B</i> gene and some major elements harbored in this region.

<p>The bold capital letter “A” represents the transcription start site (TSS), and other capital letters show different regulatory elements.</p

Expression of miR-377 in clinical HCC patients and correlation analysis with clinicopathological characteristics.

<p>(A) miR-377 was detected in 50 pairs of HCC tissues and its adjacent normal controls by quantitative RT-PCR. Data are presented as log2 of fold change of HCC tissues relative to adjacent normal regions. U6 snRNA was used as internal control. (B) Relative miR-377 expression levels in HCC tissues and adjacent normal tissues were determined by qRT-PCR. (C) The Statistical analysis of the association between miRNA level and pTNM stage (I, II, III and IV). (D) The relative expression of miR-377 in adjacent normal tissues, primary HCC tissues and lymph node metastatic tissues from 10 patients. *p<0.05, and **p<0.01, *** p<0.001.</p

MiR-377 downregulates TIAM1 through interaction with its 3’-untranslated region.

<p>(A) Schematic representation of TIAM1 3’UTR was showing the putative miR-377 target site. (B) Relative luciferase activity of the indicated TIAM1 reporter construct in HepG2 cells is shown. Firefly luciferase values were normalized to Renilla luciferase activity and plotted as relative luciferase activity. (C) RT-PCR analysis was performed to examine the effects of miR-377 on TIAM1 expression in HepG2 cells. Ectopic expression of miR-377 significantly decreased HepG2 transcripts. GAPDH was used as internal control. (D) Western blotting was performed to examine the effects of miR-377 on the expression of TIAM1. GAPDH was also detected as a loading control.</p

Localization of transcription start sites of the peanut <i>AhLEC1B</i> gene using 5′ RACE.

<p>P1 –Product of the first round PCR; P2 –Product of the second round PCR</p