Fig. 6. Microtubule association of overexpressed MIR1 point mutants. (A) The scheme shows point mutants made in the P-stretch of MIR1. MIR1A is a mutant in which all potential cdk phosphorylation sites were abolished. MIR1DE is a mutant that mimics the phosphorylated state. The scheme also shows the deletion mutants made. The N-terminal tag was either CFP, YFP, HA or 6xHis (full-length constructs only). (B) GFP-MIR1A was expressed in BHK cells during interphase or metaphase in mitosis. The cells were analyzed both for GFP fluorescence and for microtubules with a tubulin antibody. They were also stained for DNA. The arrows point to a mitotic cell showing the association of GFP-MIR1A with microtubules. (C) GFP-MIR1DE was expressed in interphase BHK cells. The cells were analyzed for GFP fluorescence and microtubule and DNA staining. The arrows point to a transfected cell showing the loss of association of GFP-MIR1DE with microtubules.

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