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. 2019 Oct 30;8:e50051. doi: 10.7554/eLife.50051

Figure 6. ALOD4 impairs hedgehog signaling by trapping accessible cholesterol.

Figure 6.

(A) ALOD4 and MβCD reduce accessible cholesterol by different mechanisms. ALOD4 binds and traps accessible cholesterol in the outer leaflet of plasma membranes of intact cells, without changing total cholesterol abundance. MβCD removes cholesterol from membranes, reducing both accessible and total cholesterol. (B) HH signaling triggered by five hours of LoSHH (5 nM) or HiSHH (30 nM) treatment following pre-treatment of cells with ALOD4 (3 or 9 µM) or MβCD (2 mM) for 1 hour. Bars denote the mean value derived from the four individual measurements shown. (C) Hmgcr mRNA levels measured with qRT-PCR after treatment with the same conditions as in (B). Bars denote the mean value derived from the eight individual measurements shown. (D) Ciliation frequency of cells after ALOD4 treatment (same conditions as in B), calculated as the number of cilia over the number of nuclei. Each point represents the ciliation frequency (derived from >30 cells) in a different imaging field. Statistical significance was determined by the Mann-Whitney test (B–D); p-values are: (B) p-value=0.0286 (all comparisons), (C) p-value=0.0002 for both comparisons, and (D) p-value=0.4857.