Pedigree and novel mutation in intron 6 c.772+27G>C of <i>ACVRL1</i> gene in the HHT family.

<p>A. Pedigree: Male (▪) and female (•) family members are shown. The filled symbols indicate patients with HHT. Dotted square indicates patient with possible HHT. B. Results of DNA sequencing of intron 6 from all members in the family and healthy individual.</p

Relative luciferase activities assay.

<p>A. A schematic diagram of the reporter constructs containing the 5′ position of intron 6 from the <i>ACVRL1</i> gene. The mutant construct containing the identified mutation site (position +27) in intron 6 of <i>ACVRL1</i> from the proband is shown, and the construct was inserted into the junction area between exon 5 and exon 6 in luciferase gene. B. Results of luciferase activity were expressed as fold increases or decreases over the empty pGL3-promoter vector (^#p<0.001). Data were from triplicate measurements with mean ± SEM.</p

Primers for amplification of introns of <i>ACVRL1</i>.

*<p>Exons amplified in each long range PCR product of <i>ACVRL1</i> gene.</p

Primers for amplification of <i>ACVRL1</i> cDNA.

*<p>Exons amplified in each RT-PCR product of <i>ACVRL1</i> gene.</p

Magnification endoscopy with narrow band imaging (NBI) pictures of the proband’s gastric mucosa.

<p>A. Example of endoscopy images showing four red spots, indicating the presence of telangiectasia of the mucosa with slight oozing of blood. B. A magnified view of a single spot on the mucosa. C. NBI of the spot shown in B.</p

The consensus Sp1 binding site.

<p>The consensus Sp1 binding site.</p

mRNA expression of the <i>ACVRL1</i>, <i>ENG</i>, and <i>VEGF</i> genes in family members with HHT.

<p>A. Expression of <i>ACVRL1</i> mRNA. The average mRNA level of healthy individuals is set to 1. The results indicate a significant decrease in <i>ACVRL1</i> mRNA levels in patients with HHT. B. Expression of <i>ENG</i> mRNA indicates a significant increase in patients with HHT. C. Assay of <i>VEGF</i> mRNA suggests a slightly increased expression in patients with HHT.</p

EMSA of the sequence at the intron 6.

<p>A. Biotin-labeled general wild-type Sp1 oligonucleotide probe was incubated with increasing amounts (2, 4, and 8 µg) nuclear extracts from PBMCs, and then Sp1 DNA binding activity was detected by EMSA. B. Biotin-SP1 wild-type probe, consisting of a double-stranded oligonucleotide containing the DNA sequence spanning nts +5 to +34 of intron 6, was incubated with increasing amounts (2, 4, and 8 µg) nuclear extracts from PBMCs, and then Sp1 DNA binding activity were assayed by EMSA. C. Biotin-Sp1 mutant probe, consisting of a double-stranded oligonucleotide containing c.772+27G>C mutation in intron 6 and sequence spanning nts +5 to +34, was incubated with increasing amounts (2, 4, and 8 µg) nuclear extracts from PBMCs, and SP1 DNA binding activity was detected by EMSA. D. 8 µg of nuclear extracts were incubated with biotin-labeled Sp1 probe (lane 2), wild-type probe (lane 4), and mutant probe (lane 6) in the presence of 100-fold excess of unlabeled Sp1 probe (lane 3), unlabeled wild–type probe (designated as wt comp, lane 5), and unlabeled mutant probe (designated as mut comp, lane 7), then SP1 DNA binding activity was determined by EMSA.</p

Two new cadinane-type sesquiterpenes from the Chinese liverwort <i>Frullania serrata</i>

<div><p>Two new cadinane-type sesquiterpenes, frullanic acid (<b>1</b>) and frullanic acid methyl ester (<b>2</b>), together with four known bibenzyls, brittonin B (<b>3</b>), 3,3′-dimethoxy-4,5-methylenedioxybibenzyl (<b>4</b>), 3,4,5,3′,4′-penlamethoxybibenzyl (<b>5</b>) and ( ± )-3-(4′-methoxybenzyl)-5,6-dimethoxyphtbalide (<b>6</b>), were isolated from the Chinese liverwort <i>Frullania serrata</i>. The structures of the new metabolites were elucidated by analysing the spectroscopic data (1D NMR, 2D NMR, HR-ESI-MS and IR). The absolute configurations of compounds <b>1</b> and <b>2</b> were determined by comparing the experimental and calculated electronic circular dichroism spectra predicted by using time-dependent density functional theory as well as the CD exciton chirality method.</p></div