Fig. 3
From: Mutations in CFAP43 and CFAP44 cause male infertility and flagellum defects in Trypanosoma and human
Immunofluorescence staining in CFAP43 and CFAP44 patients reveals an abnormal axonemal organization. a–c sperm cells from a fertile control stained with anti SPAG6 (green), a protein located in the CPC, and anti-acetylated tubulin (red) antibodies. DNA was counterstained with Hoechst 33342. c Corresponds to a and b overlay and shows that in control sperm, SPAG6 and tubulin staining superimpose. Scale bars = 5 µm. d–f SPAG6 staining is absent in sperm from the patient P43-5 homozygous for the c.2658C>T variant in CFAP43. d–i Similar IF experiments performed with sperm cells from the patient P44-2 homozygous for the c.3175C>T variant in CFAP44. Scale bar = 5 µm. Contrary to the control, the SPAG6 immunostaining (green) is abnormal with a diffuse pattern concentrated in the midpiece of the spermatozoa and is not detectable in the principle piece. j–l Sperm cells from a fertile control stained with anti RSPH1 (green), a protein of the radial spoke’s head, and anti-acetylated tubulin (red) antibodies. DNA was counterstained with Hoechst 33342. l corresponds to j and k overlay and shows that RSPH1 and tubulin staining superimpose in control sperm. Scale bar = 5 µm. m–o In sperm from the patient P43-5, RSPH1 staining (green) is significantly different from control (m) with a marked diffuse staining. p–r In sperm from the patient P44-2 the intensity of the RSPH1 staining is strongly reduced
