Fig. 1.

Experimental Design. In vitro experiments: Cortical neurons and astrocytes were cultured from C57BL/6J mice [embryonic day 18 (E18) for neurons and postnatal day 2 (P2) for astrocytes], and treated with five different concentrations of ARI, TRZ, or ARI+TRZ. CHOL, DES, 7-DHC, and LAN were analyzed by LC/MS/MS after 3, 6, and 9 days of treatment using the 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) derivatization assay. In vivo experiments: Following daily exposure to ARI (2.5 mg/kg), TRZ (10 mg/kg), ARI+TRZ (2.5 mg/kg+10 mg/kg), or vehicle (VEH) for 8 days, eight brain regions of each adult mice were dissected (n=8–9/group). Sterols, ARI, TRZ, and their main metabolites were measured by LC/MS/MS.