Figure 3

Tyrosine phosphorylation of PRC1 at Tyr-464. (a) A schematic representation of myc-PRC1-wt. Gray bars show tyrosine residue sites on PRC1. Y464 shows tyrosine residue sites mutated to phenylalanine or glutamate. (b) COS-1 cells co-transfected with the myc-PRC1-wt plus vector or Fyn were cultured for 24 h, and PP2, SU6656, or DMSO was added in the last 12 h. The immunoprecipitation of myc-PRC1-wt was performed with the anti-myc antibody. Western blotting was conducted using the indicated antibodies (c,d) COS-1 cells co-transfected with myc-PRC1-wt or myc-PRC1-Y464F plus Fyn (c) or c-Src (d) were cultured for 24 h. The immunoprecipitation of myc-PRC1-wt and the myc-PRC1-Y464F mutant was performed with the anti-myc antibody. Western blotting was conducted using the indicated antibodies.