Fig. 3: Cilium phenotyping in skeletal ciliopathy patient-derived cell lines.

A Ciliogenesis per patient, as in Fig. 2A. Patients with SRPS and CED show decreased ciliogenesis with a mean of 71 ± 13% and 80 ± 14%, respectively, in comparison to 90 ± 8% for the controls. Both patient groups show more spot cilia and more unciliated cells. Patients 5–9 with ATD and patient 4 with CEDL showed no major differences in ciliogenesis. B Cilium length of all patients, as in Fig. 2B. Patients with ATD showed longer cilia with a median of 4.81 ± 0.06 µm compared to controls (3.71 ± 0.04 µm). Patients with CED showed a shorter cilium with a median of 2.44 ± 0.05 µm, represented by a lower, thicker violin-boxplot. Patients 4 and 10, with CEDL and SRPS, respectively, showed a pear-shaped plot, in which the majority of the cilia are shorter compared to the controls, while a second population of cilia were similar to the controls. Resulting in a median of, respectively, 2.91 ± 0.10 µm and 2.30 ± 0.12 µm. C Patient IFT88 measurements, as in Fig. 2C. All patients showed an increased tip area ranging from mild to severe in comparison to the controls. Most of the patients with ATD showed a similar violin-boxplot compared to the controls, but with more outliers at the top, indicating that the majority of cilia have a normal IFT distribution, although some cilia have extremely large IFT accumulations compared to the controls. Patient 10 with SRPS also showed extremely large IFT accumualtions in a subpopulation of the measured cilia, but for this patient the majority of cilia have larger accumulations, resulting in an average tip area of 0.85 ± 0.05 µm² versus 0.43 ± 0.01 µm² for the controls. All three patients with CED showed a longer and smaller violin-boxplot, indicating that the accumulations are larger than in the controls and that there is a broad range in measurements between the cilia. A similar pattern was seen for patient 4 with CEDL.