Fig. 2

Mapping the region of RhoGDI1 for the interaction with PLK1. a Schematic representation of wild-type RhoGDI1 and truncation mutants of RhoGDI1 (1-67, 1-134, 68-204, 68-89, 90-111, 112-134 amino acids). b, c GST pull-down assay was performed with His-PLK1 expression lysates and purified GST-tagged wt RhoGDI1 or truncation mutants. The GST pull-down products were immunoblotted with PLK1 and GST antibodies. d 293 T cells were co-transfected with HA-tagged PLK1 and GFP-tagged RhoGDI1 wt or mutant (aa 90-111). Cell lysates were immunoprecipitated with a HA antibody. Immunoprecipitates and total lysates were immunoblotted with HA and GFP antibodies. e The proximity ligation assay was performed with PLK1 and RhoGDI1 antibodies in HeLa cells transfected with GFP-RhoGDI1 mutant (aa 90-111). Representative images (left) display blue (DAPI) staining nuclei and red dots (PLA signals). Scale bars: 50 μm. Quantitative data plots of PLA (right) show PLA dots per GFP-RhoGDI1 aa 90-111 positive or negative cells. **P < 0.01