Figure 12.

De novo synthesis of cholesterol and sterol precursors was accomplished by incubating hiPSC-derived neuroprogenitor cells with ${}^{13}{\mathrm{C}}_3\text{-lanosterol}$ for 24 h and monitoring for ${}^{13}{\mathrm{C}}_3\text{-sterols}$ including ${}^{13}{\mathrm{C}}_3\text{-}7\text{-dehydrocholesterol}$ and ${}^{13}{\mathrm{C}}_3\text{-cholesterol}$ (A). Simplified cholesterol biosynthetic pathway is shown (B). Neuroprogenitor cells were exposed to fenpropimorph (C–F) or spiroxamine (G–J) at $10\text{nM}$ and $\mathrm{1,000}\text{nM}$ for the same duration as the ${}^{13}{\mathrm{C}}_3\text{-lanosterol}$ incubation. Absolute values of ${}^{13}{\mathrm{C}}_3\text{-}8\text{-dehydrocholesterol}$ (C,G), ${}^{13}{\mathrm{C}}_3\text{-desmosterol}$ (D,H), ${}^{13}{\mathrm{C}}_3\text{-}7\text{-dehydrocholesterol}$ (E,I), and ${}^{13}{\mathrm{C}}_3\text{-cholesterol}$ (F,J) were detected, quantified and normalized to cell number. Three distinct differentiations were conducted for each donor and averaged; shown is the average of the three donors $\pm \text{SEM}$ ($n=3$). *$p<0.05$, **$p<0.01$, ***$p<0.001$ as determined by post hoc Dunnett’s test following one-way analysis of variance (ANOVA).