Figure 5

Interaction of KIF3B with ARHGEF10. (A) Yeast two-hybrid interaction of KIF3B with ARHGEF10. cDNAs for ARHGEF10(1–1083) and KIF3B(480–747) were subcloned into yeast two-hybrid vectors pGBKT7 and pACT2, respectively, and two-hybrid interaction was assessed by growth on the selection plate (synthetic dropout medium-Trp/-Leu/-His/-Ade/+2.5 mM 3-aminotriazole). pGBKT7-p53 and pACT2-T antigen were used as a positive control. (B) Co-localization of ARHGEF10 and KIF3B in HeLa cells. HeLa cells were transfected with an expression plasmid for FLAG-tagged KIF3B, and indicated proteins were immunostained. Arrowheads indicate centrosomes. Scale bar, 10 μm. Thin dotted lines indicate the contour of the cell. (C) Co-immunoprecipitation of KIF3B with ARHGEF10. Myc-tagged ARHGEF10 and FLAG-tagged KIF3B were expressed in COS7 cells as indicated, and association of these proteins was examined by co-immunoprecipitation. IP, immunoprecipitation; IB, immunoblotting.