Figure 6.

The DNA Binding Activity of the Mutated HD1^CCT/GHD8/OsNF-YC2 Complexes Were Measured by EMSA and ITC Assay.

(A) EMSA analysis of the interaction between DNA and HD1^CCT/GHD8/OsNF-YC2 complex containing mutated HD1^CCT (R363A, R370A, R372A, K374A, R376A, or F377A). All the mutated residues are located in the DRE region of HD1^CCT. Protein was added with increasing final concentrations of 0.25, 0.50, 1.00, and 2.00 μM in each set of four lanes (lanes 2 to 5, 6 to 9, 10 to 13, 14 to 17, 18 to 21, 22 to 25, and 26 to 29).

(B) ITC analysis of the interaction between DNA and the mutated HD1^CCT/GHD8/OsNF-YC2. The DNA sequence used for EMSA and ITC is 5′-CTC​ACT​CTC​AAC​CAC​AGC​TCG​AT-3, similar to that in Figure 2B.