Figure 4.

The TFIP11 protein binds to hPrp43 through its amino-terminus. (A) In vivo association of hPrp43 with TFIP11. Immunoprecipitation experiments were performed using anti-Flag M2 agarose from HEK293T whole-cell extracts transfected with either Flag-vector or Flag-TFIP11 and subjected to western blot analyses. The lane marked as ‘I’ contains 10% of the precipitated proteins. (B) The first 50 amino acids of TFIP11 are required for its binding to hPrp43 in vitro. Purified His-GST and His-hPrp43 proteins were immobilized on Ni-NTA beads and incubated with in vitro translated ³⁵S-labeled Flag-TFIP11 wild-type or ΔN mutant protein. Bound proteins were eluted with SDS–containing sample buffer, resolved by 12% SDS–PAGE and detected by autoradiography. An aliquot equivalent to 10% of the Flag-TFIP11 wild-type and ΔN mutant proteins used for binding was run in the lane marked ‘Input’. Molecular mass markers are indicated on the left side of the panel.