Figure 4.

In vivo phosphorylation status of Gal83. Proteins were separated by SDS-PAGE using a 1:500 bis-acrylamide:acrylamide ratio followed by α-GFP or α-HA western blot. A, Gal83 is phosphorylated in tomato protoplasts. Total protein extracts from Gal83-GFP-expressing protoplasts were treated with and without λ phosphatase (λPP) and analyzed by α-GFP western blot. The arrowheads indicate different Gal83-GFP phosphorylated forms. B, Adi3 phosphorylates Gal83 in vivo. Protoplasts expressing the indicated combinations of HA-Adi3 and Gal83-GFP were analyzed by α-GFP for analysis of the phosphorylation status of Gal83-GFP and α-HA western blot.