Fig. 4.
EFR phosphorylation site mutants fail to trigger ligand-induced phosphorylation events. Immunoblot analysis of elf18-induced phosphorylation of BAK1 (anti-BAK1-pS612) and MAP kinases (anti-p44/42) in 12-d-old seedlings expressing WT EFR and (A) EFRS753A (A#2, A#12) or EFRS753D (D#4, D#6), or (B) EFRS887A/S888A (AA#9, AA#16) or EFRS887D/S888D (DD#3, DD#8) mutants. Seedlings were treated with mock (open circles) or 1 µM elf18 (closed circles) for 15 min. Anti-GFP shows protein accumulation of WT EFR-GFP and the site-directed mutants. Panels above and below the dashed line represent immunoblots derived from replicate SDS/PAGE gels. Coomassie stained blots are shown as loading control (CBBG250). Experiments were repeated three times with similar results.