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, As biotin plays a crucial role in brain chemistry [44], and given its ability to prevent mitochondrial damage

, Stock solutions of 7b-OHC were prepared in absolute ethanol at 800 mg/ mL (2 mM), sonicated for 10 min and stored at 4 C. Biotin (Sigma-Aldrich) stock solution was prepared at 0. 5 mM in the culture medium, vortexed and then stored at À20 C. Vitamin E (a-tocopherol, ref: T3251, Sigma-Aldrich) was prepared at 80 mM in absolute ethanol (used at 400 mM), as described previously [30]. Murine oligodendrocytes 158N were seeded either in six-well plates at 30,000 cells per cm 2 (240,000 cells per well of 6 wells plates) with 2 mL of DMEM, or in Petri dishes (10-cm diameter) at 30,000 cells/cm 2 (2.35 Â 10 6 cells per Petri dish) with 10 mL of DMEM, ) and 1% antibiotics (penicillin, streptomycin) (Dutscher). 7b-hydroxycholesterol (7b-OHC) was provided by Mohammad Samadi

C. , Cells were trypsinized (0.05% trypsin-0.02% EDTA solution) and passaged twice a week, p.158

, Results are expressed as mean ± standard deviation (SD) of three independent experiments. significant difference between vehicle (ethanol) and 7b-OHCtreated cells is indicated by a (Mann-Whitney test; p < 0.05) and by b (Comparison between 7b-OHC and

. Mann-whitney, Significant difference between vehicle (ethanol) and 7b-OHC-treated cells is indicated by a (Mann-Whitney test

M. Test,

, 05). 7b-OHC: 7b-hydroxycholesterol, vol.7, p.7

, 24 h) represents 1% of the initial mass of 7b-OHC introduced in the culture medium. expressed as mean ± SD of three independent experiments. significant difference between vehicle (ethanol) and 7b-OHC-treated cells is indicated by a (Mann-Whitney test, NB: The level of 7b-OHC in 7b-OHC-treated 158N cells

M. Test, p < 0.05), and c (Comparison between vehicle (DMSO 0.05%) and biotin (10 nM) or biotin (100 nM)

M. Test, ) P MUFA: Sum of mono-unsaturated fatty acids. 7b-hydroxycholesterol, p.7

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