IOS1 Regulates Ligand-Induced FLS2/BAK1 Association.
(A) and (B) Ligand-dependent association of FLS2 to
BAK1 is reduced in the ios1-2 mutant. Col-0 or
ios1-2 seedlings were treated (+) or not (−) with
100 nM flg22 for 10 min. Total proteins (input) were subjected to
immunoprecipitation (IP) with anti-BAK1 antibodies and IgG beads followed by
immunoblot analysis using anti-FLS2 and anti-BAK1 antibodies. For
(A), Coomassie blue (CBB) is used to estimate equal loading
(bottom panel). The experiment shown in (A) is one of three
independent replicates pooled together in (B).
(C) and D) Ligand-dependent association of FLS2 to
BAK1 is augmented in the IOS1-OE3 line. Col-0 or OE3 seedlings
were treated with MgSO4 (0) or 10 or 50 nM flg22 for 10 min. Total
proteins (input) were subjected to immunoprecipitation with anti-BAK1
antibodies and IgG beads followed by immunoblot analysis using anti-FLS2 and
anti-BAK1 antibodies. The experiment shown in (C) is one of three
independent replicates pooled together in (D). For both
(B) and (D), signals were evaluated with the
ImageJ software. Values are means ± sd of three independent
biological replicates (n = 3). Different letters denote
significant difference based on a one-way ANOVA with post-hoc Tukey HSD (P <
0.05).