DDX4 during androgen manipulation.

A. Immunohistochemical localisation of DDX4 (green) in control testis. Staining is observed in late pachytene spermatocyte (PSC) cytoplasm and chromatoid body precursor structures in the perinuclear region (arrowheads). Inset shows control for the primary antibody. B. During androgen blockade (TE+Flutamide), DDX4 immunostaining intensity increased in late pachytene spermatocyte (PSC) cytoplasm. In panels A and B, cell nuclei were visualized with TOPRO (red). C. Evaluation of androgen-responsive pI isoforms of DDX4; the upper panel shows a representative image for the 2D-Western during androgen blockade (-Androgen, TE+Flutamide) compared to androgen replacement with T24 (TE+T24, +Androgen). Fourteen distinct pI isoforms were resolved. Results of the densitometric analysis of pooled samples (lower panel) from –Androgen and +Androgen groups (for details see Figure 3 legend) revealed that one isoform showed a significant (p<0.05, t-test) difference between these groups (asterix), however the other isoforms showed trends to increase or decrease with androgen replacement. Data is shown as mean ± SD (n = 3 separate experiments).