Fig. 7: CENTLEIN cooperates with SUN5 and PMFBP1 to connect the sperm tail to its head.

a Immunofluorescence analysis for CEP135 (green) and CETN1/2 (red) was performed in Centlein^+/+ and Centlein^−/− testicular germ cells. Nuclei were stained with DAPI (blue). The arrow head indicates detached CEP135 and CETN1/2 from sperm head. b, c Quantification ratio of CEP135 (b) and CETN1/2 (c) separated from the sperm head >1.5 μm in Centlein^+/+ and Centlein^−/− mice (n = 3 independent experiments). Blue dots indicate Centlein^+/+ mice and red dots indicate Centlein^−/− mice. d Ablation of Centlein impairs the localization of PMFBP1 to the coupling apparatus. Immunofluorescence analysis for PMFBP1 (green) and SUN5 (red) was performed in Centlein^+/+ and Centlein^−/− testicular germ cells. Nuclei were stained with DAPI (blue). The arrow head indicates detached PMFBP1 from sperm head. e, f Quantification ratio of SUN5 (e) and PMFBP1 (f) separated from the sperm head >1.5 μm in Centlein^+/+ and Centlein^−/− mice (n = 3 independent experiments). Blue dots indicate Centlein^+/+mice and red dots indicate Centlein^−/− mice. g Disruption of Pmfbp1 has no influence on the localization of CENTLEIN to the coupling apparatus. Immunofluorescence analysis for CENTLEIN (red) and SUN5 (green) was performed in Pmfbp1^+/+ and Pmfbp1^−/− testicular germ cells. Nuclei were stained with DAPI (blue). h Quantification ratio of CENTLEIN separated from the sperm head >1.5 μm in Pmfbp1^+/+ and Pmfbp1^−/− mice (n = 3 independent experiments). Blue dots indicate Pmfbp1^+/+mice and red dots indicate Pmfbp1^−/− mice. i Lack of Sun5 perturbs the localization of CENTLEIN to the coupling apparatus. Immunofluorescence analysis for CENTLEIN (red) and CEP135 (green) was performed in Sun5^+/+ and Sun5^−/− testicular germ cells. Nuclei were stained with DAPI (blue). The arrow head indicates detached CENTLEIN and CEP135 from the sperm head. j Quantification ratio of CENTLEIN separated from the sperm head >1.5 μm in Sun5^+/+ and Sun5^−/− mice (n = 3 independent experiments). Blue dots indicate Sun5^+/+ mice and red dots indicate Sun5^−/− mice. Data in b, c, e, f, h, j are presented as mean ± SEM. A two-tailed Student’s t test was performed, **P < 0.01.