Fig. 5.

In vitro remodeling of pre-β HDL tracers in the presence or absence of LCAT or PLTP. [¹²⁵I]TC-radiolabeled pre-β1 to -4 HDLs were incubated with plasma with or without LCAT (top panel) or PLTP (lower panel). C57Bl/6 mouse plasma was incubated with anti-mouse LCAT antiserum to immunodeplete LCAT or preimmune serum (control) (see Methods section). LCAT immunodepletion was >99% compared with preimmune incubated plasma. Plasma from wild-type (WT) or PLTP^−/− plasma was used to investigate pre-β HDL remodeling in the presence or absence of PLTP. Incubations were performed for 1 h at 4°C or for 5 min and 1 h at 37°C. Aliquots of the incubation mixtures were subjected to 4–30% NDGGE and developed with a phosphorimager. Top panel, incubations ± LCAT; bottom panel, incubations ± PLTP. 1: HMW marker; hydrodynamic size of standard proteins is shown on the left; 2: WT, 4°C, 1 h; 3: LCAT-depleted (−LCAT) or PLTP^−/−, 4°C, 1 h; 4: WT, 37°C, 5 min; 5: −LCAT or PLTP^−/−, 37°C, 5 min; 6: WT, 37°C, 1 h; 7: −LCAT or PLTP^−/−, 37°C, 1 h.