Extended Data Fig. 3: PPKs phosphorylate TZP in vitro and in vivo.
a, Identical amounts of Super:TZP-GFP and 35 S:MYC-PPK1 plasmids were transiently cotransfected into Col protoplasts. After transfection, the protoplasts were incubated in the dark for 16 h, and then treated with FR light (80 μmol m−2 s−1) for 2 h. The total proteins were extracted and treated with active (+) or inactive (boiled) or without (−) λ-PPase for 30 min, and then analyzed by immunoblots using anti-TZP, anti-MYC or anti-HSP antibodies, respectively. b, In vitro kinase assays showing that PPKs directly phosphorylate the full-length TZP proteins. CBB staining (top) and autoradiographs (bottom) are shown. c, Immunoblot assays showing the patterns of TZP proteins in Col and ppk mutant seedlings grown under FR light. Numbers below the immunoblots indicate the relative band intensities of TZP proteins normalized to those of HSP, respectively. The ratio was set to 100 for the first TZP band.
