Figure 5. SPIN6 interacts with OsRac1 in vitro and in vivo, and catalyzes OsRac1 hydrolysis in vitro.
(A) SPIN6 interacts with OsRac1 in yeast. The Mav203 yeast transformant diluted 10, 100, or 1000 times was plated on synthetic dextrose medium without Trp, Leu, or His (SD-LTH) and with 0 mM or 40 mM 3-amino-1,2,4,- triazole (3AT). EV-Empty Vector (B) The Co-IP assay of SPIN6 and OsRac1 in N. benthamiana. The GFP:Spin6 and Myc:OsRac1 plasmids were used for co-infiltration in N. benthamiana. (C) BiFC assay of SPIN6 and OsRac1 in N. benthamiana. The Spin6:CeYFP and NeYFP:OsRac1 plasmids were used for co-infiltration. (D) In vitro GAP activity assay of SPIN6. SPIN6 refers to GST-fused SPIN6 protein. OsRac1 refers to MBP-fused OsRac1 protein. GST and MBP are controls. Values are means and standard errors of three replications.