Figure 1

Mutation of plexinB1 enhances RhoD binding. A).GST-cyto-plexinB1 (WT and mutant) fusion proteins were used in GST pulldown assays with lysates of HEK293 cells expressing constitutively active (G26V) or dominant negative (T31K) RhoD-myc. B). Input of RhoD(G26V)-myc, RhoD(T31K)-myc and WT and mutant GST-cyto-plexinB1 fusion proteins. C). Expression of RhoDGTP decreases cell collapse. COS-7 cells co-transfected with plexinB1, Rnd and RhoD(G26V)-myc, or RhoD(T31K)-myc were stimulated with sema4D for 5mins, fixed and stained for plexinB1 (FITC) and myc (TRITC) and the % cell collapse scored. i).% cell collapse +/- SE, * p < 0.001 versus RhoD(T31K) 2 tailed ttest. ii) representative non-collapsed cell. iii) representative collapsed cell.