Skip to main content
. 2014 Jan 8;9(1):e84891. doi: 10.1371/journal.pone.0084891

Figure 2. The ZIM domain of TIFY8 is functional.

Figure 2

A, Y2H analysis of TIFY8 interaction with class II TIFY proteins. NINJA was included as a positive control for JAZ interaction. B, C, Analysis of TIFY8 truncations to map the interaction domain with NINJA (B) and PPD2 (C). Co-transformation of the PJ69-4A yeast strain with TIFY8 or NINJA and all TIFY family members in Gateway-compatible pGADT7 and pGBKT7 vectors, respectively. Transformed yeasts were spotted on control medium lacking Leu and Trp (-2) or selective medium additionally lacking His (-3). AD: activation domain; BD: DNA-binding domain. Controls for autoactivation are provided by transformation with the corresponding empty vector. D, Immunoblot analysis of 7 day-old Arabidopsis seedlings overexpressing the TIFY8- or JAZ1-GS fusions after 1 h treatment with either 50 µM JA or ethanol (mock). Immunoblot using the Peroxidase Anti-Peroxidase (PAP) (top) and anti-cdc2 (bottom) antibodies.