Figure 5
Specificity of anti-profilin antibodies. A) Western blot analysis of in vitro translated profilins employing antibodies directed against PFN2 (left panel), PFN3 (middle panel), and PFN4 (right panel). Note that there is no cross-reactivity of anti-PFN3 and anti-PFN4 antibodies. B) Western blot analysis of PFN2 (left panel), PFN3 (middle panel), and PFN-4 (right panel) protein expression in human testes showing varying degrees of spermatogenetic failure. Lane 1: Sertoli-cell-only appearance; lane 2: maturation arrest at meiosis; lane 3: hypospermatogenesis; lane 4: full spermatogenesis; lane 5: full spermatogenesis; lane 6: salivary gland as control. Note that PFN3 and PFN4 immunoreactive protein bands of ≈ 14 kDa were solely observed in tissue samples containing sufficient amounts of spermatids while PFN2 was detected in each tissue analysed. C) Western blot analysis of PFN1 and PFN2 (upper panels), PFN3 (middle panel), and PFN4 (lower panel) proteins in rat testis at various stages of postnatal development. Lane 1: day-15; lane 2: day-18; lane 3: day-20; lane 4: day-24; lane 5: day-26; lane 6: day-28; lane 7: day-30; lane 8: day-45; lane 9: day-60 testes. Note that PFN3 and PFN4 immunoreactive protein bands of ≈ 14 kDa were solely observed in testes at developmental stages, which contained sufficient amounts of elongating spermatids (high lightened by arrows). PFN1 and PFN2 immunoreactive bands, in comparison, were observed at all stages analysed.