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. 1999 Dec 7;96(25):14523–14528. doi: 10.1073/pnas.96.25.14523

Figure 5.

Figure 5

Cell-surface biotinylation of STEAP in transfected cells and in cancer cell lines. (A) Cell-surface biotinylation of 293T cells transfected with vector alone or with vector containing cDNA encoding 6-His-tagged STEAP. Cell lysates were immunoprecipitated with specific antibodies, transferred to a membrane, and probed with horseradish peroxidase-conjugated streptavidin. Lanes 1–4 and 6 correspond to immunoprecipitates from lysates prepared from STEAP-expressing 293T cells. Lanes 5 and 7 are immunoprecipitates from vector-transfected cells. The immunoprecipitations were performed using the following antibodies: 1, sheep nonimmune; 2, anti-Large T antigen; 3, anti-CD71 (transferrin receptor); 4, anti-His; 5, anti-His; 6, anti-STEAP; and 7, anti-STEAP. (B) Prostate cancer (LNCaP, DU145, PC-3), bladder cancer (UM-UC-3, TCCSUP), and colon cancer (LOVO, COLO) cell lines were either biotinylated (+) or not (−) before lysis. Western blots of streptavidin-purified proteins were probed with anti-STEAP antibodies. Molecular mass markers are indicated in kDa.