Figure 4.

C-Terminal Domains in TT8, EGL3, MYB75, and GL1 Are Involved in the Interactions with JAZ1 and JAZ8.

(A) and (C) Schematic diagrams show domain constructs of TT8 (A) and EGL3 (C). The diagrams display the conserved NT domain and bHLH domain. The numbers indicate positions of the first and the last amino acid of the domain constructs.

(E) and (G) Schematic diagrams show domain constructs of MYB75 (E) and GL1 (G). The diagrams display the conserved R2R3 domain and CT domain.

(B) and (D) Y2H assays to detect the interactions of JAZ1 and JAZ8 with different domains of TT8 (TT8NT and TT8CT fused with AD domain) (B) and EGL3 (EGL3NT and EGL3CT fused with AD domain) (D). pLexA vector (LexA) was used as a negative control. TTG1, which can interact with the N terminus of these bHLH factors (Payne et al., 2000), was used as a positive control.

(F) and (H) Y2H assays to test the interactions of JAZ1 and JAZ8 with different domains of MYB75 (MYB75NT and MYB75CT fused with AD domain) (F) and GL1 (GL1NT and GL1CT fused with AD domain) (H). pLexA vector (LexA) was used as a negative control. EGL3, which can interact with these MYB factors (Zimmermann et al., 2004), was used as a positive control.

(I) Expression of JAZ1, JAZ8, TTG1, and TT8NT was detected in yeast strains used for Y2H assays shown in the top panel of (B). JAZ1, JAZ8, TTG1, and LexA were detected with anti-LexA antibody (top). TT8NT was detected with anti-HA antibody (bottom).

(J) Expression of JAZ1, JAZ8, TTG1, and TT8CT was detected in yeast strains used for Y2H assays shown in the bottom panel of (B). JAZ1, JAZ8, TTG1, and LexA were detected with anti-LexA antibody (top). TT8CT was detected with anti-HA antibody (bottom).

[See online article for color version of this figure.]