Fig 2: Imaging flow cytometry analysis of AKAP4+/ASPX+/Hoechst+spermatozoa.A, 70% of motile spermatozoa permeabilized with ethanol were AKAP4+/ASPX+ positive. B, fluorescence imaging of motile spermatozoa with AKAP4 (green; Alexa 488), ASPX (red; Alexa 594), and nuclear (blue; Hoechst) staining. C and D, staining for AKAP4 (83% positive) and ASPX (71% positive) in motile spermatozoa versus secondary antibody-only controls.

Fig 3: Flow cytometry analysis of AKAP4+and ASPX+spermatozoa.A and B, normozoospermic whole semen pellets were washed, and spermatozoa were treated with 70% ethanol (A) or PBS (B) prior to staining with anti-AKAP4 and anti-ASPX primary antibodies and the corresponding secondary antibodies. Ethanol permeabilization significantly increased the fraction of AKAP4+/ASPX+ spermatozoa.