9 research outputs found
Expression of DNMTs in fetal liver.
<p>DNMT1a mRNA was determined by real time PCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044139#s4" target="_blank">Material and Methods</a> (A). Results represent the DNMT1a/β actin ratio of duplicate experimental determination of 6 different biological samples of female control (FC) and Cd-treated offspring (FT) and male control (MC) and Cd-treated offspring (MT). DNMT3a mRNA was determined by real time PCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044139#s4" target="_blank">Material and Methods</a> (B). Results are the DNMT3a/β actin ratio of duplicate experimental determination of 6 different biological samples per group; *p<0.05 compared with respective control.</p
Expression of AOX in fetal liver.
<p>AOX mRNA was determined by real time PCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044139#s4" target="_blank">Material and Methods</a> (A). Results represent the AOX/β actin ratio of duplicate experimental determination of 6 different biological samples of female control (FC) and Cd-treated offspring (FT) and male control (MC) and Cd-treated offspring (MT). Western blot analysis for AOX protein expression in liver of control (FC) and treated female (FT) and control (MC) and treated male (MT) fetus (B). AOX western blot signals were determined by densitometry and expressed as abundance ratios of AOX related to actin (C); *p<0.05 compared with respective control.</p
Liver GR1<sub>10</sub> promoter methylation and expression -relative to the control group - of offspring exposed to Cd during gestation.
<p>Liver GR1<sub>10</sub> promoter methylation and expression -relative to the control group - of offspring exposed to Cd during gestation.</p
Expression of PEPCK in fetal liver. PEPCK mRNA was determined by real time PCR as described in Material and Methods
<p>(<b>A</b>)<b>.</b> Results represent the PEPCK/β actin ratio of duplicate experimental determination of 6 different biological samples of female control (FC) and Cd-treated offspring (FT) and male control (MC) and Cd-treated offspring (MT). Western blot analysis for PEPCK protein expression in liver of control (FC) and treated female (FT) and control (MC) and treated male (MT) fetus (B). PEPCK western blot signals were determined by densitometry and expressed as abundance ratios of PEPCK related to actin (C); *p<0.05 compared with respective control.</p
Schematic diagram of the experimental protocol.
<p>The sequence of experimental procedure and number of dams and fetuses involved in each stage of the study.</p
Rat SYBR Green I assay primers.
<p>Nucleotide locations are derived from published <i>Rattus norvergicus</i>: GR mRNA (GenBank acc. no NM_012576), Phosphoenolpyruvate Carboxykinase 1 (PEPCK1) mRNA (GenBank acc. BC081900), Acyl- CoA oxidase 1 (AOX1) mRNA (GenBank acc. NM_017340), DNA methyltransferase (cytosine-5) 1 (Dnmt1) mRNA (GenBank acc. no NM_053354), DNA methyltransferase 3 (Dnmt3a) mRNA (GenBank acc. no NM_001003958) and β-actin (Actb) mRNA (GenBank acc. no NM_031144).</p
Relative position of alternative exons 1 of the rat GR gene and the CpG- rich region analyzed by bisulphite pyrosequencing at the exon 1<sub>10</sub>.
<p>The diagram is based on the previously characterized 5′-end of the rat GR gene, which contains multiple first exons. The numbering is relative to the translational start site (+1), which is located at exon 2 (A). Extent of methylation of the 9 CpG sites in the GR1<sub>10</sub> exon promoter region in fetal liver of female (B) and male (C) offspring of control and Cd-treated pregnant rats. Results were calculated by averaging the methylation level of each CpG dincleotide obtained from livers of 6 females and 6 males of different litter per group; *p≤0.05.</p
SAM and SAH concentration in fetal liver.
<p>SAM and SAH concentration in fetal liver.</p
Expression of GR in fetal liver.
<p>GR mRNA was determined by real time PCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044139#s4" target="_blank">Material and Methods</a> (A). Results represent the GR/β actin ratio of duplicate experimental determination of 6 different biological samples of female control (FC) and Cd-treated offspring (FT) and male control (MC) and Cd-treated offspring (MT). Western blot analysis for GR protein expression in the liver of control (FC) and treated female (FT) and control (MC) and treated male (MT) fetus (B). GR western blot signals were determined by densitometry and expressed as abundance ratios of GR related to actin (C); *p<0.05 compared with respective control.</p