SEPT4 Antibody - #DF13393

Figure 6. The immunofluorescence staining analysis of the presence and localization of sperm annulus ring in normal individual and FSIP2-deficient subjects. (A) The presence and localization of SEPT4 and TOMM20 staining signals in normal individual and FSIP2-deficient subjects. In normal sperm, the SEPT4 staining signal was located at the end of the MS, while the presence and localization of SEPT4 were disrupted in most of the analyzed sperm from FSIP2 deficiency subjects, including presenting on the MS, proximal region of MS, distal region of MS and absence of SEPT4 on the sperm flagella, which accompanied by MS elongation. Anti-TOMM20 (green) marked the sperm MS. Anti-SEPT4 (red) marked sperm annulus ring. The nuclei of spermatozoa were Hoechst-labeled (blue). Scale bars: 10μm. (B) Quantification of different categories of sperm annulus ring and MS defects. Immunofluorescence (IF) analysis was performed using sperm cells from three fertile controls and patients carrying FSIP2 variants (patient 1 (P1) and patient 2 (P2)). At least three semen samples from each patient were collected over 3 months through masturbation after 3-7 days of sexual abstinence. More than 200 spermatozoa from each sample were analyzed. The average sperm numbers for quantification in the control subjects and men harboring FSIP2 variants (P1 and P2) were 240, 222 and 228, respectively. Sperm annulus ring and MS defects were classified into five categories: normal MS and absent annulus (0 & 0.8%), normal MS and abnormal annulus (0 & 0.2%), long MS and abnormal annulus (38.4% & 30.4%, III-V), long MS and absent annulus (60.7% & 69.2%, VI) and long MS and normal annulus (0.9% & 0.4%, II). (C) The levels of SEPT4 in sperm cells from normal individual and FSIP2-deficient subjects analyzed by western blot. The levels of SEPT4 were reduced in men harboring FSIP2 variants. β-actin was used as internal reference.

Figure 6. The immunofluorescence staining analysis of the presence and localization of sperm annulus ring in normal individual and FSIP2-deficient subjects. (A) The presence and localization of SEPT4 and TOMM20 staining signals in normal individual and FSIP2-deficient subjects. In normal sperm, the SEPT4 staining signal was located at the end of the MS, while the presence and localization of SEPT4 were disrupted in most of the analyzed sperm from FSIP2 deficiency subjects, including presenting on the MS, proximal region of MS, distal region of MS and absence of SEPT4 on the sperm flagella, which accompanied by MS elongation. Anti-TOMM20 (green) marked the sperm MS. Anti-SEPT4 (red) marked sperm annulus ring. The nuclei of spermatozoa were Hoechst-labeled (blue). Scale bars: 10μm. (B) Quantification of different categories of sperm annulus ring and MS defects. Immunofluorescence (IF) analysis was performed using sperm cells from three fertile controls and patients carrying FSIP2 variants (patient 1 (P1) and patient 2 (P2)). At least three semen samples from each patient were collected over 3 months through masturbation after 3-7 days of sexual abstinence. More than 200 spermatozoa from each sample were analyzed. The average sperm numbers for quantification in the control subjects and men harboring FSIP2 variants (P1 and P2) were 240, 222 and 228, respectively. Sperm annulus ring and MS defects were classified into five categories: normal MS and absent annulus (0 & 0.8%), normal MS and abnormal annulus (0 & 0.2%), long MS and abnormal annulus (38.4% & 30.4%, III-V), long MS and absent annulus (60.7% & 69.2%, VI) and long MS and normal annulus (0.9% & 0.4%, II). (C) The levels of SEPT4 in sperm cells from normal individual and FSIP2-deficient subjects analyzed by western blot. The levels of SEPT4 were reduced in men harboring FSIP2 variants. β-actin was used as internal reference.