Expression of PAX8 in ovarian cancer cell lines. (A) RT-PCR was performed using total RNA from the indicated cell lines. β-actin mRNA was amplified as control. Total protein extracts prepared from the same cell lines were separated on SDS-PAGE and subjected to Western blot analysis with a specific anti-PAX8 antibody. The hybridization with actin assessed the protein uniform loading and integrity. PAX8 splice variants are indicated in both experiments. (B, C) ovarian cells were grown directly on glass coverslips and stained for indirect immunofluorescence with the anti-PAX8 antibody. The fluorescence signals were acquired at a confocal microscope, by line-wise scanning. In panel B cells are: a, medullary cells; b, cortical cells; c, SKOV-3; d, TOV-21G; e, TOV-112D; f, A2780 cells. In panel C, the extent of Pax8 silencing measured by Western blot is shown.