FIG. 1.
HPLC radiochromatograms of intracellular extracts from clone A cells following addition of 10 μM [3H]NHC for 4 h (A) and after digestion with alkaline phosphatase (B) (see Materials and Methods). Metabolites were separated by reverse-phase HPLC with a Columbus 5-μm-diameter C18 column (Phenomenex) using a Pro Star model (Varian) with manual injection. The mobile phase consisted of buffer A (25 mM ammonium acetate with 5 mM TBAP, pH 7.0) and buffer B (methanol). Elution was performed using a multistage linear gradient of buffer B.