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. Author manuscript; available in PMC: 2009 Apr 6.
Published in final edited form as: J Biol Chem. 2004 Sep 9;279(48):50230–50242. doi: 10.1074/jbc.M409130200

FIG. 8. RetSat activity in homogenized cells.

FIG. 8

Untransfected cells (solid gray trace) or Tet-induced HEKK-RetSat cells (solid black trace) were homogenized and incubated with all-trans-retinol substrate, followed by retinoid extraction and normal phase HPLC analysis. The elution profile was monitored at 290 nm for the appearance of all-trans-13,14-dihydroretinol. In control samples (short dashed black trace) cell homogenate from HEKK-RetSat cells was boiled 10 min at 95 °C prior to incubation with substrate. The addition of 0.4 mM NADH or NADPH had no effect on the yield of all-trans-13,14-dihydroretinol. The experiment was performed in duplicate. The peak numbers represent 13-cis-retinol (1), all-trans-13,14-dihydroretinol (2), 9,13-di-cis-retinol (3), 9-cis-retinol (4), and all-trans-retinol (5). mAU, milliabsorbance units.