Figure 9.

OsDIS1 promotion of OsNek6 degradation in vivo. A, OsNek6 degradation assay. The Agrobacterium strains carrying the DsRed-OsDIS1 and GFP-OsNek6 plasmids were coinfiltrated into N. benthamiana, with the DsRed and DsRed-OsDIS1(H71Y) plasmids as controls. The HA-GFP plasmid was used as an internal control for protein synthesis. Three days after infiltration, samples were collected for protein and RNA extraction. B, Inhibition of OsNek6 degradation by MG132. A 50 μm solution of MG132 was infiltrated into N. benthamiana 24 h before sampling with DMSO as a control. The extracted protein and RNA were used for western-blot and RT-PCR analyses, respectively.