RIN4 and NDR1 Interact in Planta.
Coimmunoprecipitation of RIN4 and NDR1 in N. benthamiana and Arabidopsis.
(A) Transient coexpression in wild-type N. benthamiana plants. Immunoblot of anti-HA and anti-T7 immunoprecipitated proteins isolated 40 h after inoculation from wild-type N. benthamiana leaves hand-infiltrated with Agrobacterium strains expressing HA:NDR1 and T7:RIN4. Total protein extracts were immunoprecipitated with anti-HA (NDR1) and anti-T7 (RIN4) antibodies. Immunoprecipitated proteins were detected by immunoblotting anti-T7 (top panel) and anti-HA (bottom panel) immunoblots. Protein sizes are indicated at the left side of the immunoblots.
(B) Expression and coimmunoprecipitation of HA:NDR1 and native RIN4 in Arabidopsis. Total protein extracts were immunoprecipitated with either anti-HA (positive control) or anti-RIN4 antibodies, followed by protein gel blot analysis with anti-HA. “Beads only” indicates a negative control whereby total extracts were incubated at 4°C in the absence of antibody.