Figure 3.

PPM1G physically interacts with p27 in cells and in vitro. A. PPM1G interacts with p27 in 293T cells. GFP-p27 and Flag-PPM1G (WT or DN) were co-transfected into 293T cells. PPM1G was immunoprecipitated (IP) with anti-Flag antibody and the presence of p27 in the IP complex was determined by Western blot. B. PPM1G interacts with p27 in vitro. In vitro translated Flag-p27 was incubated with GFP or PPM1G purified from E. coli as GST fusion protein in an in vitro binding buffer. GST pull-down product was then analyzed by Western blot using p27 antibody.