Additional file 1.

Validation of TLC/phosphorimaging approach for determining cholesterol and 24,25EC synthesis. (C) CHO-7 cells were incubated with [1-¹⁴C]-acetate for 24 h under conditions of partial OSC inhibition to yield comparable levels of cholesterol and 24,25EC. Samples were pooled and neutral lipids extracted. Varying amounts of the extracts were applied to a thin-layer chromatography plate and the greatest amount was assigned an arbitrary relative concentration of 1. After development, bands corresponding to authentic cholesterol and 24,25EC were visualized by phosphorimager and quantified by densitometry (A, B). Alternatively, bands were cut, eluted and quantified by scintillation counting (D, E). Equations for the lines of best fit (A) y = 1.01x; R² = 0.99; (B) y = 1.00x; R² = 0.99; (D) y = 1.00x; R² = 0.99; (E) y = 1.00x; R² = 0.99.

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Wong et al. Lipids in Health and Disease 2007 6:10   doi:10.1186/1476-511X-6-10