Figure 2. NbMIP1.1a interacts with ToMV MP and Tm-2² in vivo.

Firefly luciferase complementation imaging assays for in vivo interaction of NbMIP1.1a with ToMV MP (A) and Tm-2² (B). Panels show luminescence images of N. benthamiana leaves agro-infiltrated with nLUC-NbMIP1.1a and ToMV MP-cLUC or Tm-2²-cLUC. The combinations of nLUC-NbMIP1.1a and cLUC, nLUC and ToMV MP-cLUC, nLUC and Tm-2²-cLUC were included as negative controls. (C) NbMIP1.1a co-immunoprecipitated (co-IP) with ToMV MP and Tm-2². NbMIP1.1a-Myc was co-expressed with ToMV MP-HA or Tm-2²-HA in N. benthamiana leaves by agroinfiltration. NbMIP1.1a-Myc co-expressed with HA-nLUC was introduced as a negative control. At 48 hours post infiltration (hpi), leaf lysates were immunoprecipitated with anti-HA beads, then the immunoprecipitates were assessed by western blotting using anti-Myc (upper panel) and anti-HA antibodies (middle panel). In addition to immunoblotting for co-IP, presence of NbMIP1.1a-Myc, ToMV MP-HA, Tm-2²-HA and HA-nLUC in the cell lysates were also analyzed (lower panel).