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. 2020 Nov 2;11:5520. doi: 10.1038/s41467-020-19113-0

Fig. 4. Cfap45−/− mice replicate a motile ciliopathy phenotype.

Fig. 4

a–f Motile cilia of the LRO have a “9 + 0” axoneme (a). Cfap45 is detectable in the mouse embryonic LRO by in situ hybridization (antisense probe) (b); control probe (sense) shows no hybridization (c). Black scale bar equals 100 µm. d–f CFAP45 (red) localizes to motile LRO cilia (d, e) but not non-motile (primary) cilia (f) of the LRO by IFM. g–i Compared to wild-type control, Cfap45−/− mice show LRA defects including situs inversus totalis with heart malpositioned on the right side (g). h Rotational speed of motile LRO cilia is significantly reduced in Cfap45−/− embryos (n = 74 cilia from four embryos) compared to control (n = 87 cilia from 4 embryos); data are mean ± SEM, significance assessed by two-tailed t test (***p < 0.0001). See also Supplementary Videos 3 and 4. i Particle image velocimetry (PIV) analysis shows significantly reduced fluid flow at the LRO of Cfap45−/− embryos compared to control. Yellow and red color indicate leftward flow, blue indicates rightward flow. j–p Similar to respiratory cilia, sperm flagella have a “9 + 2” axoneme (j). In contrast to control (k), Cfap45−/− sperm flagellar waveforms show reduced curvature and amplitude under conditions inducing hyperactivation (l). m, n CFAP45 (red) is detectable by IFM in testis cryosections of heterozygous littermates (m) but not Cfap45−/− males (n). o Compared to control, motility of Cfap45−/− sperm is significantly reduced (n = 195 sperm over three independent experiments for each; data are mean ± SEM; significance assessed by two-tailed t test, ***p = 0.0003). p Compared to Cfap45+/- littermates, Cfap45−/− sperm show significantly reduced curvilinear velocity (n = 99 or 70 sperm over three independent experiments for Cfap45+/− and Cfap45−/−, respectively; data are mean ± SEM; significance assessed by two-tailed t test, ***p = 0.0005). See also Supplementary Videos 5 and 6. Ciliary and flagellar axonemes (green) are detected using anti-acetylated α tubulin (AcTub) antibody. Merge images include Hoechst stain (blue) to indicate nuclei. White scale bars equal 10 µm. n = 6 images from 2 experiments (b, c, m, n); n = 4 images from one experiment in d, f.