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. 2013 Jul 25;9(7):e1003653. doi: 10.1371/journal.pgen.1003653

Figure 1. JAZ proteins interact with bHLH3, bHLH13, bHLH14 and bHLH17.

Figure 1

(A) Yeast two-hybrid (Y2H) assay to detect interactions of JAZs with bHLH3, bHLH13, bHLH14 and bHLH17 factors. Twelve Arabidopsis JAZs were fused with the LexA DNA binding domain (BD) in pLexA respectively. The bHLH3, bHLH13, bHLH14 and bHLH17 were fused with the activation domain (AD) in pB42AD respectively. Interactions (represented by blue color) were assessed on 2% Gal/1% raffinose/SD/-Ura/-His/-Trp/-Leu/X-β-Gal medium. (B) Bimolecular fluorescence complementation (BiFC) assay to detect interactions of JAZ1 and JAZ10 (fused with nYFP) with bHLH3, bHLH13, bHLH14 and bHLH17 (fused with cYFP). Construct pairs were coexpressed in leaves of N. benthamiana. YFP fluorescence was detected 50 hours after infiltration. The nuclei were indicated by DAPI (4,6-diamidino-2-phenylindole dihydrochloride) staining. (C) Y2H assay to test interactions of bHLH13 domain constructs with twelve JAZs. The schematic diagram shows the bHLH13 domain constructs. The conserved JID domain and bHLH domain were shown with blue and gray box respectively. The numbers indicate the positions of amino acid. Different bHLH13 domains were fused with AD in pB42AD, and JAZs were fused with BD in pLexA. (D) Y2H assay to test interactions between different domains of JAZ8 and JAZ11 with bHLH3, bHLH13, bHLH14 and bHLH17 respectively. The schematic diagram shows the JAZ8 and JAZ11 domain constructs. The conserved ZIM and Jas domains are indicated by gray and green boxes respectively. Different domains of JAZ8 and JAZ11 were fused with BD, and these bHLH factors were fused with AD individually.