Skip to main content
. Author manuscript; available in PMC: 2012 Dec 20.
Published in final edited form as: Biochemistry. 2011 Nov 29;50(50):10860–10875. doi: 10.1021/bi201264y

Figure 2. Analysis of multimerization of rat SR-BI.

Figure 2

A, CHO cells transiently transfected with rat SR-BI-V5 plasmid for 36h were lyzed in lysis buffer containing 2% SDS, NaPFO or Triton X-100. After heating samples with or without 50 mM DTT at 95°C for 2 min, suitable aliquots (15 μg protein) were separated on 8% PAGE and immunoblotted with rabbit anti-peptide (C-terminal peptide) rat SR-BI antibody as described under “Materials and Methods.” B, Highly purified double-membraned plasma membrane preparations from luteinized rat ovaries were solubilized in lysis buffer containing 2% SDS, heated at 95°C for 2 min in the presence and absence of 50 mM DTT and suitable aliquots subjected to 10% PAGE. Following transfer, membranes were immunoblotted with rabbit anti-peptide (C-terminal peptide) rat SR-BI antibody probed with rabbit anti-peptide (C-terminal peptide) rat SR-BI antibody (20) followed by a horseradish peroxidase-conjugated goat anti-rabbit IgG (Sigma). The bands were visualized using ECL Plus™ Western blotting detection reagents (GE Healthcare, Piscataway, NJ) as described under “Materials and Methods.” Arrows indicate the bands corresponding to monomers, dimer and teramer forms of SR-BI. The blot shows that rat SR-BI dimers or oligomers are resistant to types and the presence of detergent, but sensitive to sulfhydryl reducing agent, dithiothreitol.