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. Author manuscript; available in PMC: 2010 Sep 2.
Published in final edited form as: Biochem J. 2009 May 13;420(2):305–315. doi: 10.1042/BJ20081909

Figure 2.

Figure 2

Sterol content and biosynthesis in J774 cells adapted to grow in a cholesterol-free medium. (A) Cholesterol and desmosterol content of cells at different times of growth after the complete replacement of RPMI containing 10% FBS with either RPMI containing 10% LPDS or DCCM-1. (B and C) HPLC profiles of radioactivity (B) and absorbance at 206 nm (C) of sterols from cells at day 21 of growth in DCCM-1. DCCM-1 was supplemented with 40 μCi of [14C]acetate for 3 days and the cell extracts containing the nonsaponifiable lipid fraction were analyzed by reverse-phase HPLC and in-line radioactivity counting. The arrows indicate the elution times of desmosterol (des), cholesterol (chol) and ergosterol (erg). d, days; m, months; is, internal standard; nd, not detected. (D) Average cholesterol and desmosterol levels in cells maintained in RPMI containing 10% FBS or at days 21 to 32 of growth in DCCM-1. Bars show the mean and whiskers indicate the SEM of data from seven separate cell cultures.