Sf9 cells were transfected with a VHDL-receptor cDNA expression vector, as described in Materials and Methods. Transfected and wild-type cells (control) were lysed, and the presence of VHDL receptor analysed in the growth medium and the cell lysate.
Panel A: Dot blots of 3 µg of cell lysate (lysate) or concentrated growth medium (medium) were spotted onto PVDF membrane, and processed as described in Materials and Methods.
The Western blot was probed with anti-VHDL-receptor antiserum (1:6,000, secondary antibody 1:10,000; Western). Ligand blots were carried out with biotinylated VHDL (1: 8,000, Ligand Biotin-VHDL) or FITC-labeled VHDL (1:8000, Ligand FITC-VHDL)
Panel B: Ligand blot with FITC-VDHL following electrophoresis. Left lane: purified receptor (0.1 µg), right lane: protein concentrated from the Sf9-lysate (3 µg).