Figure 1

Expression pattern of nucleolin in spermatogenic cells. A) FACS profile of total testicular cells (TTC), meiotic spermatocytes; (4n); and spermatid cells (n) purified from rat testis. B) Real time PCR analysis of nucleolin mRNA normalized with respect to Naca1. Differences (n-fold) in expression of nucleolin were compared across gametic diploid cells, meiotic spermatocytes and spermatid cells keeping low expressing cells as the reference. Each value plotted represents an average of 3 independent experiments ± standard deviation. 2n, gametic diploid cells (testicular cells from 10 day old rat); 4n, meiotic spermatocytes; and n, spermatids. C) Western blot analysis of nucleolin using mouse monoclonal nucleolin antibody using goat anti-mouse IgG HRP conjugate as secondary antibody. 100 μg of protein was loaded in each lane. 2n, diploid cells; 4n, meiotic spermatocytes; and n spermatid cells. D) Immunolocalisation of nucleolin using mouse monoclonal nucleolin antibody, followed by counterstaining with goat anti-mouse Alexa 568 and 488. Nucleus was stained with DAPI (1 μg/ml). HeLa cells, (panel a-b); gametic diploid cells, (panel c-d); meiotic spermatocytes, (panel e-f); and spermatid cells, (panel g-h).