Fig. 1
From: The DELLA proteins interact with MYB21 and MYB24 to regulate filament elongation in Arabidopsis
Interactions of DELLAs with MYB21 and MYB24. a Schematic diagram of MYB21 and MYB24 domain constructs. The conserved R2R3 domain, and the NYWG/SM/VDDI/LWS/P motif are indicated by yellow and blue respectively. The numbers indicate positions of the first and the last amino acid of the domain constructs. b Yeast Two-Hybrid (Y2H) assay to detect interactions of MYB21NT and MYB24NT with DELLAs and their derivatives. MYB21NT and MYB24NT were individually fused with the LexA DNA binding domain (BD) in pLexA. DELLAs and their derivatives were individually fused with the activation domain (AD) in pB42AD. Interactions of MYB21NT and MYB24NT with the AD domain in pB42AD empty vector were used as negative controls. Interactions (represented by blue color) were assessed on 2% Gal/1% raffinose/SD/−Ura/−His/−Trp/−Leu/X-β-Gal medium. c Schematic diagram of DELLAs domain constructs. The R part of DELLAs contain the conserved DELLA domain (green). The numbers indicate positions of the first and the last amino acid of the domain constructs. d Y2H assay to detect interactions of R domains of DELLAs with MYB21, MYB24, and their derivatives. R domains of DELLAs were individually fused with BD domain in pLexA. MYB21, MYB24 and their derivatives were individually fused with the AD domain in pB42AD. Interactions of R domains of DELLAs with the AD domain in pB42AD empty vector were used as negative controls. Interactions were assessed on 2% Gal/1% raffinose/SD/−Ura/−His/−Trp/−Leu/X-β-Gal medium. e-g In vitro pull-down assay to detect the interactions of RGA (e), RGL1 (f), RGL2 (g) with MYB21 and MYB24. Purified MBP, MBP-MYB21, and MBP-MYB24 fusion proteins were incubated with the TAP-RGA, TAP-RGL1, or TAP-RGL2 expressed in TAP-RGA, TAP-RGL1, or TAP-RGL2 transgenic Arabidopsis plants. Bound proteins were washed, separated on SDS-PAGE, and immunoblotted with the anti-c-myc antibody (α-myc). The input lane shows the expression level of TAP-RGA, TAP-RGL1, or TAP-RGL2 expressed in TAP-RGA, TAP-RGL1, or TAP-RGL2 transgenic plants. The positions of purified MBP, MBP-MYB21 and MBP-MYB24 on SDS-PAGE are indicated with asterisks (stained by Coomassie blue). The original data can be viewed from Additional file 4: Figure S3a-d