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. 2002 Jan;22(1):298–308. doi: 10.1128/MCB.22.1.298-308.2002

FIG. 9.

FIG. 9.

Physical interaction between τ131 and B”. Interaction between coexpressed τ131 and B” was revealed by copurification assay. High Five cells were coinfected with three different combinations of two recombinant baculoviruses, one expressing a GST-fused subunit (GST-τ131 or GST-Gea1) and the other expressing 8His-B” or calmodulin-TBP. Extracts were prepared as described in Materials and Methods. A glutathione purification was performed, and 50 μl of eluted fractions obtained were loaded onto SDS-PAGE gels. (A) Coomassie blue staining of an 8% polyacrylamide gel. Lanes 1 and 5 are molecular mass markers; some masses are indicated on the left (in kilodaltons). Combinations of recombinant baculoviruses are indicated at the tops of lanes 2, 3, and 4. Lane 6 is a control showing 8His-B” purified to homogeneity (heparine hyper-D and IMAC resin). The positions of B” are indicated with dots in lanes 2 and 6. (B) Input (10 mg of crude extracts) and bound proteins after glutathione purification (50 μl of eluted fractions) were analyzed by Western blotting. The membrane was probed with polyclonal antibodies directed against 8His-B” and TBP as indicated on the left. Lane 7 is the same control as that shown in lane 6 of panel A.