Fig. 4.

AtD14 interacts with MAX2 independently of SMXL6/7/8. The in vivo interactions between HA-AtD14 and GFP–MAX2 revealed by co-immunoprecipitation (Co-IP) assay in protoplasts prepared from the smxl6 smxl7 smxl8 triple mutant (A) and the wild-type Col-0 (B). After transformation and incubation for 11 h, protoplasts were pre-treated with rac-GR24 for 1 h, cells were broken, and then immunoprecipitation (IP) using agarose-conjugated anti-GFP monoclonal antibody was carried out in the presence or absence of 100 µM rac-GR24. The HA-AtD14 recombinant protein was detected by anti-HA monoclonal antibody, and the GFP–MAX2 fusion protein and GFP were detected by anti-GFP monoclonal antibody. Input means extracted crude proteins without immunoprecipitation.