Figure 3

Western blot analyses of YN- or YC-fused NH proteins and RH proteins in protoplasts. (A) Rice protoplasts were transfected individually with an YN-NH plasmid plus an YC-RH plasmid as described in Figure 1. The white “*” symbols indicate cleaved products of NH1, NH2, and NH3, respectively. (B) Protoplasts were transfected with an YC-NH plasmid and an YN-RH plasmid. The negative control contained plasmids pSY736 and pSY735 encoding YN and YC respectively. Protein samples were prepared as in Figure 1 and gel loading was normalized to GUS activity. The NC membranes were probed with α-Myc then α-HA, or α-HA then α-Myc, sequentially. Molecular weight: NRR = 14.2kD; RH1 = 19.2kD; RH2 = 18.1kD; and RH3 = 18.3kD. Molecular weights of YFPN, YFPC, NH1, NH2, NH3, NH4, and NH5 are listed in Figure 1. Each protein is marked with a white arrowhead.