Fig. 8. The ABD-containing N-terminal regions of Bpag1 can selfassociate but require the presence of the unique regions upstream from the ABD to do so. (A) Immunoblots showing 1/10 input of the lysates used for performing immunoprecipitations. GFP-Nterm1 and FLAG-Nterm1, GFP-Nterm2 and FLAG-Nterm2, or GFP-Nterm2 and FLAG-ABD were co-expressed in Cos-1 cells. All of the GFP- and FLAG-tagged proteins were expressed relatively evenly, with the exception of the FLAG-Nterm2, which was very highly expressed. This was typical of other results looking at the expression of these proteins (data not shown). (B) Immunoprecipitation with an anti-FLAG antibody pulls down GFP-Nterm1 (lane 2) in cells coexpressing FLAG-Nterm1 and GFP-Nterm1, and GFP-Nterm2 (lane 4) in cells co-expressing FLAG-Nterm2 and GFP-Nterm2. However, the GFP-Nterm2 was not co-immunoprecipitated with the FLAG-ABD fusion protein (lane 6). Lanes 1, 3, and 5 are immunoprecipitations corresponding to lanes 2, 4 and 6, respectively, which were performed without the anti-FLAG antibody as controls for the experiment.

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