![Figure 3. The BH1 domain mediates the interaction between anti-Bcl-2-scFv-TAT and Bcl-2. (A) Binding of anti-Bcl-2-scFv-TAT to intracellular Bcl-2. Bcl-2 was immunoprecipitated from RBL cells with anti-Bcl-2 immobilized on protein A-agarose. The binding of anti-Bcl-2-scFv-TAT (lane 1) or nonspecific scFv-TAT (lane 2) to intracellular Bcl-2 was determined by Western blot analysis using antibody against the myc tag on the scFv. (B) The sequence of the Bcl-2-BH domains and their homology to other BH domains in the Bcl-2-family proteins. The box encloses the BH domains, and the Bcl-2 sequences before and after the box complete the fragments for PCR. (C) Bcl-2 fragments BH1, BH2, BH3, and BH4 were labeled with [35S]-methionine in vitro and added to anti-Bcl-2-scFv-TAT immobilized on agarose beads. Retained [35S]-labeled fragments were determined by SDS-PAGE and autoradiography. One representative experiment of 3 is shown.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/102/7/10.1182_blood-2002-12-3921/6/h81935002003.jpeg?Expires=1726825256&Signature=QHdmzq9pL16fJUOLCe2Spn9tWFTvArN1Q1GJ17A97VHyN8ihRTluWfyL9OccdVGA4KKO7n31~raMXqbUTsPbWAje5KknnensUP-LGMgaaeailMQd3ep0Kj5hKffXlf2pvNCEj2YzSCKKa3~1vAexR0Z3p7z5Euys7qaCo90~-ekZVLXowB5loKcRm6tQ0AXe5c4SVWFBNkcn0vos7taKyvu85xsuiTXb~5T6GJnxkLU-gGmM8o1AtDQ77nzrVscdGQfEmLHvyq-SjVogS6T5eHtafCx6CzBCtwzmPEllZjU8DrAI6jafsWL7BPUWGethogjkJ8x7qpXkhp55sQZeKw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
The BH1 domain mediates the interaction between anti-Bcl-2-scFv-TAT and Bcl-2. (A) Binding of anti-Bcl-2-scFv-TAT to intracellular Bcl-2. Bcl-2 was immunoprecipitated from RBL cells with anti-Bcl-2 immobilized on protein A-agarose. The binding of anti-Bcl-2-scFv-TAT (lane 1) or nonspecific scFv-TAT (lane 2) to intracellular Bcl-2 was determined by Western blot analysis using antibody against the myc tag on the scFv. (B) The sequence of the Bcl-2-BH domains and their homology to other BH domains in the Bcl-2-family proteins. The box encloses the BH domains, and the Bcl-2 sequences before and after the box complete the fragments for PCR. (C) Bcl-2 fragments BH1, BH2, BH3, and BH4 were labeled with [35S]-methionine in vitro and added to anti-Bcl-2-scFv-TAT immobilized on agarose beads. Retained [35S]-labeled fragments were determined by SDS-PAGE and autoradiography. One representative experiment of 3 is shown.
