Fig. 6

Fig. 6

a IFM analysis of primary cilia in control and patient dermal fibroblasts (HDFs) cultured in the presence of serum. Primary cilia (arrows) are marked with anti-ARL13B (green) and anti-acetylated α-tubulin (Ac-tub, red), and nuclei are stained with DAPI (blue). Scale bars, 40 µm. Scale bar in zoomed image, 2 µm. b Quantification of ciliary frequency in control and patient HDFs serum depleted for 0–72 h (control ≥ 132 cells per time point, patient ≥125 cells per time point, data average of n = 3 independent experiments; 0 h: P = 0.002, 24 h: P = 0.268, 48 h: P = 0.349, 72 h: P = 0.397). c Quantification of ciliary length in control and patient HDFs serum depleted for 0–72 h (control ≥ 63 cells per time point, patient ≥ 82 cells per time point, data average of n = 4 independent experiments; 0 h: P = 0.507, 24 h: P = 0.053, 48 h: P = 0.440, 72 h: P = 0.169). d Overview of ciliary lengths in control HDFs (upper panels) and patient HDFs (lower panels) subjected to serum depleted for 48 h followed by serum addition for 0–24 h. Cilia are marked with anti-ARL13B (arrows, red) and centrosomes (ciliary basal bodies) are marked with anti-PCTN-2 (asterisks, white). Scale bar, 2 µm. e Quantification of cilia frequency based on data presented in d (control ≥ 131 cells per time point, patient ≥ 142 cells per time point, data average of n = 3 independent experiments; 6 h: P = 0.480, 12 h: P = 0.176, 16 h: P = 0.070, 20 h: P = 0.039, 24 h: P = 0.002). f Quantification of cilia length based on data presented in d (control ≥ 60 cells per time point; patient ≥ 60 cells per time point, data average of n = 3 independent experiments; 6 h: P = 0.479, 12 h: P = 0.908, 16 h: P = 0.444, 20 h: P = 0.001, 24 h: P = 0.0004)). Data are represented as mean ± SD and significance was determined using an unpaired, two-tailed Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, n.s.: not significant.