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5 and a Pearson correlation significance with p value were selected for further analysis. The selected genes were then inferred by applying the ARACNe algorithm. To maximize the statistical significance, we referred to a large dataset of human normal and neoplastic B-cells as well as human B-cell lines that has been reported previously [45, 48] and is available at GEO datasets ""type"":""entrez-geo"",""attrs"":""text"":""GSE2350"",""term_id"":""2350""GSE2350 and ""type"":""entrez-geo"",""attrs"":""text"":""GSE12195"",""term_id"":""12195""GSE12195 ARACNe was performed using geWorkbench software, with bootstrapping, at a p value threshold of sample (referred to as ��NF-��B signature��), and finally (iii) we divided the analyzed samples into two groups (NF-��B+ and NF-��B?) based on whether the expression levels of the NF-��B signature were above or below the median value. 2.3. Quantitative RG7204 in vivo RT-PCR IFI16 mRNA expression analysis was performed through quantitative real-time RT-PCR (qRT-PCR) on RNA extracted from peripheral blood purified B-cell subsets achieved from three healthy donors (n = 3, two men and one woman, age between 32 and 36 years). Total RNA was extracted from purified B-cell subsets using the High Pure RNA isolation kit (Roche, Mannheim, Germany) and stored at ?80��C. Then, total RNA was reverse transcribed and amplified employing the Quantitect SYBR Green RT-PCR kit (Qiagen, Hilden, Germany) following the manufacturer's procedure. In brief, SYBR green real-time PCR assay was performed in a 20?��L PCR mixture volume consisting of 10?��L of 2X Quantitect SYBR green RT-PCR Master Mix (Qiagen), containing HotStarTaq DNA polymerase, 400?nM of each oligonucleotide primer, 0.2?��L of 100X QuantiTect RT Mix (containing Omniscript and Sensiscript reverse transcriptases), and 100?ng of extracted RNA. The IFI16 primer sequences [53] were as follows: IFI16F: 5��-ACAAACCCGAGAAACAATGACC-3��, nt 1464�C1485, (RNArefseq ""type"":""entrez-nucleotide"",""attrs"":""text"":""NM_005531.2"",""term_id"":""112789561"",""term_text"":""NM_005531.2""NM_005531.2) and IFI16R: 5��-GCATCTGAGGAGTCCGAAGA-3�� nt 1584�C1565, (RNArefseq {""type"":""entrez-nucleotide"",""attrs"":{""text"":""NM_005531.2"",""term_id"":""112789561"",""term_text"":""NM_005531.