Figure 5

Comparative analyses of normal and subfertile mice using MAST antibody. (A) Comparative Western blotting with testes and sperm lysates from normal (XY) and subfertile (XYq-del) mice probed with MAST antibody shows presence of a 20 kDa protein in comparable quantities in the testis lysates of both. MAST was absent in the subfertile sperm proteome. Tubulin antibody served as the loading control. (B) Shows cytoplasmic localization of the protein in the entire cell types of normal (XY) testis. In the subfertile (Yq-del) testis there is a substantial reduction in signal, indicating either reduced expression or alternatively lack of localization to the correct locus. Pre-immune serum was used as negative control and PI was the counterstain. Magnification - 100× and scale bar = 25 μm. (C) Comparative immunolocalization in sperms: Immunolocalization with antibody to MAST protein showed that in normal (XY) sperms the protein localizes to the acrosome and sperm tail. In sperms from subfertile (XYq-del) mice a very faint signal is observed only in the midpiece of sperm tail. DAPI was used as the counterstain for nucleus and pseudo color red was used for representation. Magnification - 63×; scale bar = 10 μm.