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. 2015 Jun 1;10(6):e0128315. doi: 10.1371/journal.pone.0128315

Fig 1. PAX8 directly modulates NRP2 gene expression.

Fig 1

(A) FRTL-5 cells were transfected with a siRNA that specifically targets rat PAX8. PAX8 and Neuropilin-2 (NRP2) expression levels were measured on total RNA by qRT-PCR 24h, 48h and 72h after siRNA transfection. The values are means ± SD of three independent experiments in duplicate, normalized by the expression of β-actin and expressed as fold change with respect to the untransfected FRTL-5 cells, whose value was set at 1.0. Statistical analysis uses t test (p ≤ 0.05). (B) Chromatin extracted from cross-linked FRTL-5 cells was immunoprecipitated using in parallel an unrelated antibody (anti-tubulin) or an antibody against PAX8. The immunoprecipitates were analyzed by PCR with oligonucleotides corresponding to the rat NRP2 promoter region. Parallel PCR were performed with total input DNA obtained from unprecipitated aliquot of similarly treated chromatin sample and from no template control (NTC). Schematic representation of the upstream region of the rat NRP2 gene. PAX8-binding site is represented as a striped box.