Figure 4.

The activated Tm-2² resides at the PM. A, HR cell death is induced by Tm-2² in the presence of TMV MP and autoactive MHD mutant D481V, but not by wild-type Tm-2² alone in N. benthamiana. Cell death was visualized by trypan blue staining (lower panel) at 48 hpi. Solid line circles indicate cell death; dashed line circles indicate no obvious cell death. B, Cell fractionation assays show that activated Tm-2² is associated with the membrane. Soluble (S) and microsomal membrane (M) fractions were separated by ultracentrifugation. C, Aqueous two-phase partitioning assays show that the activated Tm-2² is partitioned in the PM phase. Upper phase (U) and lower phase (L) were obtained by aqueous two-phase partitioning of microsomal membrane fractions from B.