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. 2000 Apr 11;97(8):4363–4368. doi: 10.1073/pnas.97.8.4363

Figure 5.

Figure 5

PBP interactions with RAR and ER. (A) GST pulldown assay demonstrating ligand-dependent interaction between PBP–YFP and GST–RAR or GST–ER LBD. (B and C) Emission ratio detection of FRET from CFP–RAR full-length (B) or CFP–RAR LBD (C) to PBP–YFP on addition of RA or the synthetic RAR ligand TTNPB. The figures on the right of the panels show the percentage of FRET efficiency calculated by using the YFP-photobleach protocol at the end of each experiment. (D) GST pulldown assay demonstrating ligand-dependent interaction between PBP–YFP and GST–ER LBD. (E) Emission ratio monitoring of the interaction between CFP–ER LBD and PBP–YFP. Tamoxifen reduces the interaction, and estrogen promotes it. (F) The fluorescence intensity of FRET and donor channels for the above experiment is shown. The artifact at around 400 s represents refocusing, which cancels out when the YFP and CFP are ratioed. The CFP fluorescence after photobleaching YFP is Fd.