Fig. 4

The localization and levels of flagella-associated proteins in ARMC2 patients and controls. (A-C) Immunofluorescence assays of flagella-associated proteins, including TOMM20 (mitochondrial outer membrane protein), SPAG6 (a component of the CP complex), and IFT 88 (a component of IFT-B subcomplex) in ARMC2 patients and controls. Anti-TOMM20 (red in A) localized at mitochondrial sheath, Anti-SPAG6 (red in B), Anti-IFT-88 (green in C) were normally localized along the sperm flagella in the control sperm. A complete absence of SPAG6 signals was observed in P2. Anti-Ac-Tubulin (green in A and B, red in C) marked the sperm flagella, and Hoechst (blue) marked the nucleus of spermatozoa. Scale bar: 10 μm. (D) WB assays analyzed the expression levels of TOMM20, SPAG6, and IFI-88 in sperm obtained from ARMC2 individuals and normal controls. The results of WB assays were consistent with those of immunofluorescence assays described above. β-Actin was used as a loading control