Fig. 1: Loss of HSD17B4 leads to dysgenesis of primary cilia in different cell types.

a, b Primary cilia were visualized in fibroblasts derived from healthy controls (Cont #1, #2) and HSD17B4-deficient patients (c.422_423DelAG; Patient #1, #2) by immunostaining with an anti-ARL13B (green) antibody. Nuclei were counterstained with Hoechst 33342 dye (blue). Data are presented as the mean ± SEM (n = 3, independent biological replicates). P-value vs. healthy controls determined by one-way ANOVA followed by Tukey’s multiple comparisons test [Ciliated cells: Cont #1 vs. Patient #1 p = 0.0008, vs. Patient #2 p = 0.0002; Cont #2 vs. Patient #1 p = 0.0005, vs. Patient #2 p = 0.0001; Cilium length: Cont #1, 2 vs. Patient #1, 2 p < 0.0001]. c HSD17B4 expression was abrogated in HSD17B4 patient-derived fibroblasts. Western blot analysis was performed using the indicated antibodies. Representative blot from three independent experiments is presented d, e RPE cells were transfected with either scrambled (Sc) siRNA or siRNA targeting HSD17B4 (siHSD17B4) for 72 h. Primary cilia were immunostained with an anti-acetylated-α-tubulin antibody (red), and nuclei were counterstained with Hoechst 33342 dye (blue). f, g HSD17B4-knockout (KO) and wild-type (WT) SH-SY5Y/Cas9 cells were immunostained with an anti-ARL13B (green) antibody and Hoechst 33342 (blue) dye. The number of ciliated cells and the ciliary length of the cells were quantified using a fluorescence microscope. Scale bar, 5 μm. Data are presented as the mean ± SEM (n = 3, independent biological replicates). P-value vs. Sc non-targeting siRNA treatment or WT cells determined by two-tailed unpaired t-test [Ciliated cells: Sc vs. siHSD17B4 p = 0.0032 (e), WT vs. KO p = 0.0160 (g); Cilium length: Sc vs. siHSD17B4, WT vs. KO p < 0.001 (e, g)]. h Primary cilia-mediated signaling was assessed by western blotting using the indicated antibodies. Experiments were repeated three times. Source data are provided as a Source Data file.