Pro-neural transcription factor expression is increased in more invasive/metastatic prostate cancer cell-lines. (a) The expression of pro-neural and neuroendocrine transcripts was measured by quantitative RT-PCR using four different cell lines. In the bicalutamide resistant cells (black bars), the androgen-independent cells C4-2 (blue bars) and the androgen-independent metastatic C4-2b cells (green bars) the expression of Nts, Ascl1 and Hes6 with metastatic potential of the lines. The cell line data is presented in order of metastatic potential in a xenograft model of prostate cancer (LNCaP on the left-hand side having limited metastatic potential progressing through to C4-2b with the potential to generate bone metastases upon implantation). Data is represented as fold change as comnpared with the parental LNCaP cell line (light blue bars with asterisk). Also there is expression of NeuroD1 and Neurogenin2 (Ngn2) in the metastatic cells but to a lesser degree. No changes in the expression of Hes1 were observed in any of the cell lines. All the data was normalized using the average of four housekeeping genes (Gapdh, Sdh, Ubc and Tbp) and the ratio against the parental line (purple bars) was plotted. A table with the Ct values obtained for each of the genes prior to analysis is provided (b) Immunohistochemistry for Ascl1 on paraffin embedded sections from LNCaP and bicalutamide-resistant LNCaP cell-lines.