Figure 1.

Mip1 interacts with hMps1 in vitro and in cell lysates. (A) Recombinant full length Mip1 binds hMps1 in vitro. Equivalent volumes of glutathione-bead bound GST-hMps1-ha, GST-hMps1-KD-ha or GST alone were added to E. coli lysates from cells expressing 6X-histidine tagged Mip1 or empty vector. The beads were collected, washed and analyzed by SDS-PAGE and immunoblot with anti-6X histidine and anti-GST antibodies. (B) Mip1 and hMps1 co-precipitate from cell lysates. Antibodies towards hMps1 or Mip1 were used to immunoprecipitate proteins from cycling U2OS cells and precipitated material was probed for the presence of Mip1 and hMps1 using either anti-Mip1 or anti-hMps1 antibodies. Multiple bands in Mip1 containing lanes are degradation products.