Figure 3. Tm-22 (VAALLA) mutant, NbMIP1.1a and ToMV MP could exist in the same complex in plants.
(A) The Tm-22 (VAALLA) mutant did not induce HR when co-expressed with ToMV MP-Myc (upper), and failed to induce resistance against TMV-GFP (lower). Tm-22-HA or Tm-22 (VAALLA)-HA were agroinfiltrated into wild-type N. benthamiana leaves with either ToMV MP-Myc (upper) or TMV-GFP (lower). Photos were taken at 4 dpi under UV light. (B) Tm-22 (VAALLA)-HA co-immunoprecipitated with both ToMV MP and NbMIP1.1a (top). YFP-NbMIP1.1a co-immunoprecipitated with both ToMV MP and Tm-22 (VAALLA) mutant (middle). Loading controls were also analyzed by western blot (bottom). Tm-22 (VAALLA)-HA and YFP-NbMIP1.1a were co-expressed with ToMV MP-Myc or empty Myc vector in N. benthamiana leaves through agroinfiltration. At 60 hpi, leaf lysates were immunoprecipitated with anti-HA or anti-GFP beads, then the immunoprecipitates were assessed by western blotting using anti-HA, anti-Myc or anti-GFP antibodies as indicated.