FIGURE 7: Immunoblot analysis.
(A) PAP expression in yeast cells expressing AtBI-1 or AtBI-1DC.
(B) AtBI-1 or AtBI-1DC expression in yeast cells expressing PAP.
Yeast transformants were grown in SD-U-L glucose media. The expression of PAP and AtBI-1 was induced for indicated hours, lysed and fractionated into membrane (P18) and cytosolic (S18) components. The amount of 10 mg protein was separated on 15 % SDS-PAGE. Proteins were transferred into nitrocellulose membrane and probed with polyclonal PAP antiserum and monoclonal V5 antibody. The membrane marker Dpm1p and the cytosolic marker Pgk1p were used as controls to show equal amount of loading and lack of cross-contamination.