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. 2007 Oct 15;27(24):8815–8823. doi: 10.1128/MCB.01085-07

FIG. 2.

FIG. 2.

AIRE binds CycT1 in vitro and in cells. (A) AIRE binds the GST.CycT1 chimera in vitro. Binding reactions were performed between GST and the GST.CycT1 chimera, which were expressed in Escherichia coli, and AIRE, which was expressed in HeLa-MAGI cells. (Left) Lanes 1 to 4 contain specific pulldowns, and lane 6 contains 10% of the input AIRE protein. (Right) Input of GST proteins, which were visualized by Coomassie blue staining of SDS-PAGE. Molecular size markers (in kilodaltons) are given in lane M. (B) AIRE interacts with the endogenous CycT1 and Cdk9 proteins in 1C6 cells. The Flag epitope-tagged AIRE protein (lanes 1 to 6) was expressed in 1C6 cells. Total cell lysates were immunoprecipitated (IP) with αFlag M2 agarose beads (lane 2), αCycT1 (lane 5), or mouse IgG (lanes 3 and 6) antibodies and examined for the presence of CycT1, Cdk9, Cdk7, and AIRE by Western blotting (WB) with αCycT1, αCdk9, αCdk7, and αFlag antibodies, respectively. Lanes 1 and 4 represent 10% of the input of indicated proteins.