Skip to main content
. 2018 Nov 26;8:17335. doi: 10.1038/s41598-018-35614-x

Figure 5.

Figure 5

A patient-derived iPSC line containing the CEP290-R1746Q variant (#725) exhibits altered cilia morphology and function as well as diminished response to Shh-N stimulation. (A) Maximum intensity projections of confocal image z-stacks of iPSC cultures typically used for assessment of ciliated cell fractions and cilium length measurements. Immunostaining was used for detection of ciliary axonemes (Arl13b) and for subcellular localisation of CEP290. White squares indicate zoomed-in regions. White arrows point to localisation of CEP290 at the base of cilia. Scale bars = 10 μm and 5 μm. (B) Quantification of cilia-bearing cell fractions in iPSC cultures, n = 36 z-stacks/cell line; three independent experiments. (C) Cumulative frequency distribution of cilia lengths in iPSC cultures, n = 748 (#574 line), 539 (#725 line), 822 (#110 line); three separate experiments. ****p < 0.0001, Kolmogorov-Smirnov t-test. (D–H) FRAP curves generated as described in Fig. 4. Bar graphs representing the recovered fraction for each fluorescent protein analysed are inlayed in respective curve plots. n ≥ 23 for Smo-EGFP, n ≥ 25 for Arl13B-mCherry, n ≥ 15 for 5HT6-EGFP, n ≥ 18 for SSTR3-EGFP, and n ≥ 16 for IA-EGFP. n = cells per category, three independent experiments. Data are represented as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, one-way ANOVA post hoc Holm-Sidak’s multiple comparisons test.