Fig. 2

A missense homozygous variant in the ZCWPW1 gene was identified in this infertile patient. A Family pedigree of the patient with ZCWPW1 mutation. The proband is represented by the black square. B Sanger sequencing confirmed the mutation of c.1064C > T) in this family. The black arrow denotes the mutation position C The precise locations of the variant and changed amino acids in ZCWPW1. ZCWPW1 contains two domains: CW-TYPE zinc finger domain (amino acid positions 250–304) domain and PWWP (amino acid positions 317–383) domain. (D) Multiple sequence alignment of the ZCWPW1 protein for different species. The black arrow denotes the position of the variant (p. P355L). E Western blot reflected a marked decline in Mut-ZWCPW1 expression compared with WT-ZCWPW1. Vector is negative control. F The grayscale analysis of the protein bands was shown. Data represent the mean ± SD from three independent experiments. Student’s t-test, *P < 0.05, **P < 0.01. G The immunofluorescence staining showed the expression of ZCWPW1 in the spermatozoa from a fertile control individual and the patient. (Scale bars = 5 μm). H Effects of ZCWPW1 mutation on the structural conformation of ZCWPW1 protein. Left panel is the structural conformation of ZCWPW1 protein wild type and right is the mutant. The amino acid positions are labeled in red. The structures of the ZCWPW1 protein were changed by the mutation of c.1064 C > T [p.P355L]