Figure 1.

GRFs interact with PBL19 and other RLCKs. A) GRF6 interacts with PBL19 in Arabidopsis protoplasts. Col-0 protoplasts expressing the indicated constructs were treated with chitin or flg22 or neither, and Co-IP assays were performed and detected with anti-FLAG and anti-HA immunoblots. B) GRF6 interacts with PBL19 in vitro. His-tagged PBL19 was incubated with either GST-tagged GRF6 recombinant protein or GST. GST pull-down assays were performed, and input and immunoprecipitated proteins were detected using anti-GST and anti-His antibodies. C) GRF6 interacts with multiple RLCK VII members. The indicated Nluc and Cluc constructs were transiently expressed in N. benthamiana for luciferase complementation assays. Data are presented as mean relative luminescence units (RLUs) ± se. Different letters indicate significant differences at P < 0.05 (n = 12, one-way ANOVA, Tukey post-test). CPK5 was used as a negative control. D) Co-IP assays showing that BIK1 interacts with GRF6 in vivo. Ten-day-old seedlings were treated with or without flg22 and then samples were harvested. Total proteins were extracted and subsequently incubated with anti-IgG Affinity Gel and the HA antibody. The total and precipitated proteins were examined by immunoblotting using antibodies against BIK1 and HA, respectively. Each experiment was repeated at least 3 times with similar results, and results from one representative experiment are shown.