Fig. 3: Acetate supplementation restores primary ciliogenesis disrupted by HSD17B4 mutations.

a–c Fibroblasts derived from healthy controls (Cont #1, Cont #2) and HSD17B4-deficient patients (422_423DelAG; Patient #1, #2) were treated with sodium acetate (10 mM) for 24 h. a Intracellular acetyl-CoA levels were assessed using an acetyl-CoA ELISA kit from whole-cell lysates. Representative data from three independent experiments (n = 3) are presented as the mean ± SEM. [Patient #1, #2: Cont vs. Acetate p < 0.001]. b, c Primary cilia were immunostained with an anti-ARL13B (green) antibody, and the nuclei were counterstained with Hoechst 33342 (blue) dye. Data are presented as the mean ± SEM (n = 3, independent biological replicates). [Ciliated cells: Cont vs. Acetate; Cont #1 p = 0.0016, Cont #2 p = 0.0006, Patient #1 p = 0.0018, Patient #2 p = 0.0011; Cilium length: Cont vs. Acetate; Cont #1, 2, Patient #1, 2 p < 0.001]. d Schematic representation of HSD17B4 domains: dehydrogenase (pink), hydratase (blue), and SCP2 (yellow). The patient-derived mutations (G16S and N457Y) are indicated. e–g SY5Y/Cas9 HSD17B4-KO cells were transfected with hemagglutinin (HA)-tagged WT or mutant (G16S or N457Y) HSD17B4 constructs. e The expression of HSD17B4 was examined by western blotting. Representative blot from three independent experiments is shown. f, g HSD17B4 WT or mutant-overexpressing SY5Y/Cas9 HSD17B4-KO cells were co-stained with an anti-HA (red), anti-ARL13B (green) for primary cilia, and Hoechst 33342 (blue) for nuclei. Data are presented as the mean ± SEM (n = 3, independent biological replicates). [Ciliated cells: Cont vs. Acetate; Cont p = 0.0013, WT p = 0.0004, G16S p < 0.0001, N457Y p < 0.0001; Cilium length: Cont vs. Acetate; Cont p < 0.0001, WT p = 0.0016, G16S p < 0.0001, N457Y p < 0.0001]. P-value vs. between the indicated groups determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Scale bar, 5 μm. Source data are provided as a Source Data file.