Fig. 6
Physical association between LecRK-VI.2 and BAK1. a In vitro maltose-binding protein (MBP) pull-down assay of LecRK-VI.2KD interaction with BAK1KD. Recombinant MBP or MBP-LecRK-VI.2KD was incubated with GST-BAK1KD and pulled down with amylose resin beads. Input and bead-bound proteins were analyzed by immunoblotting with monoclonal anti-MBP and anti-GST antibodies. b Co-immunoprecipitation (Co-IP) analysis of LecRK-VI.2-FLAG association with BAK1-GFP in N. benthamiana. Total proteins (input) of N. benthamiana leaves transiently co-expressing LecRK-VI.2-FLAG and BAK1-GFP, or transiently expressing BAK1-GFP alone, were immunoprecipitated with anti-FLAG affinity agarose beads and the precipitates were analyzed by immunoblotting with monoclonal anti-FLAG and anti-GFP antibodies. The leaves were treated with (+) or without (−) 0.8 mm NADP+ for 10 min. c Co-IP analysis of LecRK-VI.2-HA association with BAK1-GFP in Arabidopsis. Total proteins of Arabidopsis leaves from the BAK1:BAK1-GFP/LecRK-VI.2:LecRK-VI.2-HA transgenic plants were immunoprecipitated with monoclonal anti-GFP antibody/protein G plus agarose and the precipitates were analyzed by immunoblotting with polyclonal anti-GFP and monoclonal anti-HA antibodies. The leaves were treated with (+) or without (−) 0.8 mm NADP+ for 10 min
