Trypan blue staining of SOBIR1 variants expressed in N. benthamiana
(A)AtSOBIR1 and various mutants fused to C-terminally enhanced green fluorescent protein were transiently expressed in N. benthamiana by Agrobacterium-mediated transient expression (agroinfiltrations). Pictures of representative leaves stained with trypan blue exhibit levels of cell death. All assays were performed three times, and a representative photograph is shown. Scale bar, 2 cm.
(B) All infiltrated N. benthamiana leaves were collected after phenotype analysis, mixed with 2× loading buffer (v/v), and boiled at 95°C for 15 min. Expression levels of all proteins were then checked by western blotting with anti-GFP antibody.