X chromosome inactivation analysis.

<p>After digestion with or without Hpa II, DNA was used to amplify a polymorphic CAG repeat within the androgen receptor (AR) gene. The size of the allele is determined by the number of repeats. The area under the peak indicates the degree of amplification of that allele. II5 and II3 inherited the allele 276 from their mother (I 1) and allele 265 from their father, II7 inherited the other allele 286 from I 1. After digestion with Hpa II, their paternal allele was predominantly amplified and represented the inactive. In contrast, III2 inherited the allele 276 from her mother (II3), which was predominantly amplified and represented the inactive. Thus four of them showed skewing of XCI (XCI ratio≥80∶20).</p

Biochemical findings^a.

a<p>VLCFAs normal ranges: C26 0.378–0.642 µmol/L; C24/C22 0.606–1.16; C26/C22 0.011–0.023.</p>b<p>unit: µmol/L.</p

XCI pattern of X-ALD carriers, related noncarriers and normal controls.

<p>XCI pattern of X-ALD carriers, related noncarriers and normal controls.</p

T cloning findings.

<p>T cloning findings.</p

Alignment of ALDP proteins in different species.

<p>The black bar indicates the position of p.283H in the ALDP sequence.</p

Rapid Decolorization of Phenolic Azo Dyes by Immobilized Laccase with Fe₃O₄/SiO₂ Nanoparticles as Support

Fe₃O₄/SiO₂ nanoparticles with particle size below 30 nm were used as the support for laccase immobilization through glutaraldehyde coupling. Investigation of the immobilized laccase was carried out by X-ray diffractometry (XRD), transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), vibrating sample magnetometry (VSM), UV–vis spectrophotometry, and cyclic voltammogram (CV) measurements. Two phenolic azo dyes, Procion Red MX-5B and azophloxine, were selected to investigate the enzyme activity of the immobilized laccase toward degradation of phenolic azo dyes. The immobilized laccase presents unusual performance for dye decolorization and easy separation with an external magnetic field. Finally, the possible mechanism for the unusual decolorization of phenolic azo dyes by the immobilized laccase is discussed

ISL-1 promotes pancreatic islet cell proliferation by forming an ISL-1/Set7/9/PDX-1 complex

<p>Islet-1 (ISL-1), a LIM-homeodomain transcription factor, has been recently found to be essential for promoting postnatal pancreatic islet proliferation. However, the detailed mechanism has not yet been elucidated. In the present study, we investigated the mechanism by which ISL-1 promotes β-cell proliferation through regulation of CyclinD1 in HIT-T15 and NIT-1 cells, as well in rat islet mass. Our results provide the evidence that ISL-1 promotes adult pancreatic islet β-cell proliferation by activating CyclinD1 transcription through cooperation with Set7/9 and PDX-1 to form an ISL-1/Set7/9/PDX-1 complex. This complex functions in an ISL-1-dependent manner, with Set7/9 functioning not only as a histone methyltransferase, which increases the histone H3K4 tri-methylation of the CyclinD1 promoter region, but also an adaptor to bridge ISL-1 and PDX-1, while PDX-1 functions as a RNA pol II binding modulator. Furthermore, the formation of the ISL-1/Set7/9/PDX-1 complex is positively associated with insulin-like growth factor-1 treatment in NIT and HIT-T15 cells <i>in vitro</i>, while may be negatively correlated with age <i>in vivo</i>.</p

Expression patterns of <i>Dlx3</i> and EMP genes at the mRNA level during amelogenesis.

<p>Real-time RT-PCR was used to analyze the expression levels of <i>Dlx3</i> and EMP genes using RNA extracted from tooth germs at the indicated stages. The expression level at PN1 was set at 1, and the fold-changes at other stages were calculated relative to PN1. The data represent three independent experiments, and are shown as mean ± SD. *<i>P</i> <0.05 <i>vs</i>. the control (PN1 group). <i>Enam</i>, Enamelin; <i>Amelx</i>, Amelogenin; <i>Ambn</i>, ameloblastin; <i>Tuft</i>, Tuftelin-1; <i>Mmp20</i>, Matrix metalloproteinase 20; <i>Klk4</i>, kallikrein 4; <i>Odam</i>, Odontogenic ameloblast-associated protein; <i>Amtn</i>, Amelotin.</p

TDO mutation of DLX3 inhibits the activation of EMP genes by wild-type DLX3.

<p>(A) The plasmids pCI-neo (control empty vector, con), V5DLX3^WT, and V5DLX3^TDO were transfected into LS8 cells. Equal amounts of V5DLX3^WT or V5DLX3^TDO plasmid were added to each group, and the total amounts of plasmid in the groups were kept constant using pCI-neo. Over-expression of V5DLX3^WT or/and V5DLX3^TDO were analyzed by western blot using antibody against V5-tag. A representative figure from three independent experiments is shown. (B) After over-expression, the mRNA expression levels of <i>Enam</i>, <i>Amelx</i>, <i>Klk4</i>, and <i>Odam</i> were evaluated. Values were obtained from three independent experiments, and are presented as mean ± SD. *<i>P</i> <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. (C) Left panel: western blot for the protein expression levels of ENAM, AMELX, and KLK4 after transfection (representative of three independent experiments). Right panel: densitometric analysis of images of 3 independent experiments. *<i>P</i> <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. (D) After co-transfection with pCI-neo (empty control vector), V5DLX3^WT, V5DLX3^TDO, or both V5DLX3^WT and V5DLX3^TDO plasmids, the relative transcriptional activity of the 5 reporter constructs containing potential DLX3 response elements were analyzed by luciferase assays. Data were compared with the control group (con), and are presented as mean% ± SD. *<i>P</i> <0.05 between the DLX3^WT group and the DLX3^WT+DLX3^TDO group. <i>pEnam-E1</i> represents pGL<i>Enam-E1</i>, etc.</p

Regulation of <i>Enam</i>, <i>Amelx</i>, <i>Klk4</i>, and <i>Odam</i> by DLX3-overexpression and knockdown.

<p>(A) Over-expression of DLX3 was determined by real-time RT-PCR and western blot. Values are mean ± SD of the data from three independent experiments. *<i>P</i> <0.05 <i>vs</i>. control (con) group. (B) After 36 h of transfection, the expression of EMP genes was assessed. The mRNA expression of <i>Enam</i>, <i>Amelx</i>, <i>Klk4</i>, and <i>Odam</i> were significantly up-regulated. Values are presented as mean ± SD. *<i>P</i> <0.05 <i>vs</i>. the control (con) group. (C) After 48 h of transfection, elevated protein expression of ENAM, AMELX, and KLK4 were detected by western blot. Upper panel: western blot bands of ENAM, AMELX, and KLK4 (representative of three independent experiments). Lower panel: densitometric analysis of images of 3 independent experiments. *<i>P</i> <0.05 <i>vs</i>. the control (con) group. (D) Expression levels of DLX3 were determined after transfection with 3 independent <i>Dlx3</i>-specific small interfering RNAs (<i>Dlx3</i> siRNA #1, #2, #3), or a nonspecific siRNA, NS siRNA. Values are from three independent experiments, and shown as mean ± SD. *<i>P</i> <0.05 <i>vs</i>. NS group. (E) After DLX3 silencing (using <i>Dlx3</i> siRNA #3), the expression of EMP genes was assessed. The expression of <i>Enam</i>, <i>Amelx</i>, <i>Klk4</i>, and <i>Odam</i> was significantly down-regulated, and then successfully rescued by transfection of plasmid pCI-neo-V5DLX3^WT. Values are mean ± SD of data from three independent experiments. *<i>P</i> <0.05. (F) At the protein level, expression of ENAM, AMELX, and KLK4 were also down-regulated by DLX3-knockdown and then rescued by pCI-neo-V5DLX3^WT transfection, as analyzed by western blot. Upper panel: western blot bands of ENAM, AMELX, and KLK4 (representative of three independent experiments). Lower panel: densitometric analysis of images of 3 independent experiments. *<i>P</i> <0.05.</p