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. 2014 Mar 27;165(1):319–334. doi: 10.1104/pp.114.237891

Figure 7.

Figure 7.

Phosphorylation and interaction of HSFA4A with MAP kinases. A, Phosphorylation of HSFA4A substrate with immunoprecipitated MPK3 and MPK6 kinases analyzed by in-gel kinase assay. Anti-MPK3 and anti-MPK6 antibodies (Sigma) partially recognize both kinases. PLANT indicates extracts obtained from wild-type Arabidopsis (Landsberg erecta). Control indicates gel without HSFA4A substrate. B, In vitro phosphorylation of purified HSFA4A by MPK3 and MPK6. MyelinBP was used as artificial substrate in positive control reaction. His-tagged MPK3/MPK6 was used in phosphorylation reactions. HSFA4A was tagged with maltose-binding protein (MBP-HSFA4). C, Interaction of HSFA4A with MPK3 and MPK6 in Y2H system. A indicates proteins fused to the activation domain, B indicates proteins fused to the DNA-binding domain, and Ctr indicates empty vector with the DNA-binding domain. D, Interaction of HSFA4A-nYFP, MPK6-cYFP, and MPK3-cYFP in A. tumefaciens-transformed tobacco leaf cells. BIFC indicates interacting proteins. Bars = 20 μm.