Figure 3.

OsDSK2a Interacts with the Gibberellin Metabolism Regulator EUI.

(A) Interactions of full-length and truncated EUI proteins with OsDSK2a in yeast. At left, a schematic diagram of full-length and truncated EUI fusion proteins with GAL4-BD is shown; at right panel, the growth of transformed yeast cells on SD/-Trp-Leu and SD/-Trp-Leu-His medium, respectively. SD/-Trp-Leu (X-gal) shows a filter assay for β-galactosidase activity of the transformants. pGBK-EUI/pGAD-T7 and pGBK-T7/pGAD-OsDSK2a combinations were used as negative controls.

(B) Coimmunoprecipitation assays to detect the interaction of OsDSK2a with EUI in vivo. Total proteins extracted from callus cotransformed with EUI-HA/ OsDSK2a-GFP or VTC1-HA /OsDSK2a-GFP were immunoprecipitated with anti-HA antibodies, washed four times (W1–W4), and the eluted fractions (Elute) separated by SDS-PAGE before immunoblotting with anti-GFP antibodies.