Fig 2: Individuals carrying RSPH1 mutations retain markers of the radial spoke stalk and the inner and outer dynein arms (ODA). High-resolution immunofluorescence analysis in respiratory epithelial cells obtained by nasal biopsy is shown in unaffected controls compared with individuals carrying mutations in RSPH1 with anti-acetylated-α-tubulin used as a marker to stain the entire axoneme (red) compared with a marker of the radial spoke stalk complex, anti-ROPN1L (green) and antisera against the ODA protein DNAH5 and the inner dynein arm (IDA) protein DNALI1 (green). Nuclei are DAPI-stained to show the DNA (blue). (A) ROPN1L, (B) DNAH5 and (C) DNALI1 immunostaining in PCD-166 II:1 is unaltered compared with healthy individuals. Scale bars represent 10 μm.

Fig 3: Immunofluorescence of nasal ciliated cells revealed the absence of DNAH5 and DNALI1 of proband 1. Anti-acetylated tubulin monoclonal antibody was used to mark the ciliary axoneme. DNAH5 was used to label the outer dynein arm (ODA), DNALI1 was used to label the inner dynein arm (IDA) and DAPI was used to label the nuclei.

Fig 4: Pedigree of the family with PCD and the clinical features of the proband from this family. (A) There is one patient (the proband) in the family with PCD. Circles indicate to females. Squares indicate males. Solid symbols indicate patients. Half solid symbols indicate carriers of the identified mutations. The arrow indicates the proband. (B) Chest CT scan of the proband showed complete situs inversus, bronchiectasis and Recurrent bronchitis and pneumonia. (C) The transmission electron micrograph (TEM) of bronchial ciliary epithelium from the proband. The red arrow and the green arrow represent IDAs and ODAs, the black arrow indicates absence of IDAs and ODAs (A–C). A schematic diagram of the bronchial ciliary epithelium cross-section(D). (D–F) Immunofluorescence of bronchial ciliary epithelium revealed the existence of DNAH5, DNAH9 and DNALI1 of proband. Anti-acetylated tubulin monoclonal antibody was used to mark the ciliary axoneme. DNAH5 and DNAH9 were used to label the outer dynein arm (ODA), DNALI1 was used to label the inner dynein arm (IDA).

Fig 5: Immunofluorescence staining of spermatozoa from a control individual and the proband (II:1) with dynein arm protein antibody and α‐tubulin. Anti‐α‐tubulin was used to label the spermatozoa axoneme (green). Spermatozoa were counterstained with DAPI (blue) as a nuclei marker. (A) Outer dynein arm heavy chain protein DNAH5 (red) and α‐tubulin (green) localized along the entire sperm from the control, but complete absence of DNAH5 from the spermatozoa axoneme from the proband. (B) Inner dynein arm light intermediate chain protein DNALI1 (red) was detected along the entire sperm with α‐tubulin (green) from the control, but was partly detected in the spermatozoa axoneme from the proband. Scale bars: 2 μm.