Chromatin inmunoprecipitation (ChIP) assays.

<p>(A) The PCR amplified regions comprising the predicted NFκB binding sites (kBS1, kBS2 and kBS3) are indicated as rectangles, with location of these specific DNA segments indicated by numbers. (B) TPC and CGTH cells were treated with TNF-α and ChIP assays performed using IgG (mock) and NFκBp65 antibodies. Imnmunoprecipitated DNA was analyzed by qPCR with primers targeted to the predicted NFκB binding sites (kBS1, kBS2 and kBS3). The quantitative data reflect occupancy calculated as % input and represented as mean±SD of two independent experiments. *: P<0.05; **: P<0.01.</p

Identification of functional NF-κB responsive elements in the 5’-flanking region of the human <i>TBX15</i> gene.

<p>(A) Location and sequence of the predicted NF-κB binding sites (kBS1, kBS2 and kBS3) in the studied region of the <i>TBX15</i> promoter, with indication of altered sequence in mutated reporter constructs (underlined). Site directed mutagenesis was performed using P2 reporter construct to obtain the mutated reporter constructs. (B) Luciferase analysis of each mutated reporter constructs. <i>Left panel</i>, co-tansfected with the pCDNA3-p65 expression vector (p65) or the pCDNA3 empty vector (EV). <i>Right panel</i>, in nonstimulated or stimulated TNF-α cells. At least three independent experiments were performed with duplicates. Bars represent the mean±SD. * indicates p-value < 0.05 and ** p-value < 0.01.</p

Expression of <i>TBX15</i>mRNA and <i>CXCL1</i>mRNA in BHT cell lines after stimulation with TNF-α and PMA/I.

<p>(A) qRT-PCR of <i>TBX15</i>mRNA analysis in BHT and BHT IkBαSR cells after 3h of TNF-α or PMA/I treatment. (B) qRT-PCR of <i>CXCL1</i>mRNA analysis in BHT and BHT IkBαSR cells after 3h of TNF-α or PMA/I treatment. Data were normalized to <i>RPL27</i> and expressed as fold change referred to untreated cells whose <i>TBX15</i>mRNA or <i>CXCL1</i>mRNA relative expression was defined as 1. Data are mean ± SD of mRNA levels in triplicates. ** indicates p-value < 0.01.</p

Expression of <i>TBX15</i>mRNA in different cell lines after stimulation with TNF-α.

<p>(A) qRT-PCR of <i>TBX15</i>mRNA analysis in FRO, CGTH, TPC-1 and HeLa cells after 2h of TNF-α treatment. (B) qRT-PCR of <i>TBX15</i>mRNA (left) and <i>CXCL1</i>mRNA (right) analysis in p65^+/+ and p65^<i>-/-</i> MEF cells after TNF-α treatment. Data were normalized to <i>RPL27</i> and expressed as fold change referred to untreated cells whose <i>TBX15</i>mRNA or <i>CXCL1</i>mRNA relative expression was defined as 1. Data are mean±SD of mRNA levels of two independent experiments in triplicates. ** indicates p-value < 0.01.</p

Differences for after-irradiation DNA damage, according to <i>WDR3</i> haplotypes.

<p>Global haplotype association P-value: 0.79.</p

Correlation between DNA damage values (spontaneous <i>vs</i> after irradiation).

<p>Correlation between DNA damage values (spontaneous <i>vs</i> after irradiation).</p

Spontaneous and after irradiation BNMN frequencies, according to gender and histological DTC type.

a<p>Wilcoxon signed rank test.</p>b<p>Student's t-tests.</p

Differences for basal DNA damage, according to <i>WDR3</i> haplotypes.

<p>Global haplotype association P-value: 0.31.</p

Differences for after-irradiation DNA damage, according to <i>WDR3</i> polymorphisms.

<p>SE: standard error; CI: confidence interval.</p>*<p>Adjusted by gender, age and diagnosis.</p><p>C, D: Codominant and Dominant models, respectively.</p

Differences for basal DNA damage (BNMN), according to <i>WDR3</i> polymorphisms.

<p>SE: standard error; CI: confidence interval.</p>*<p>Adjusted by gender, age and diagnosis.</p