SL Destabilizes the D14 Protein.
(A) to (F)
GUS histochemical activity of D14pro:D14:GUS plants in Col-0
(A) or max2-1
(B) background after 24-h treatment with mock (left) or GR24 (right). Fluorescence image of young CaMV35Spro:D14:GFP seedlings after 24-h treatment with mock (C) or GR24 (D). Fluorescence image of D14pro:D14:GFP seedlings after 24-h treatment with mock (E) or GR24 (F). Bars = 500 μm.
(G) D14:GFP response to GR24 analyzed by immunoblot using α-GFP antibody in D14pro:D14:GUS plants in Col-0 (wild type) or max2-1.
(H) Time course of D14:GFP accumulation in response to GR24. Protein extracts (10 μg) from 5-d-old D14pro:D14:GFP seedlings treated with GR24 (+) or mock (−) for 0 to 24 h were separated by 10% SDS-PAGE and identified as a 56.5-kD band.
(I) After a 24-h GR24 treatment, seedlings were maintained in MS with (+) or without GR24 (−) for 1 to 8 h.
(J) Dose response of D14:GFP to 24-h treatments with increasingly higher concentrations of GR24.
(K) D14:GFP response to GR24 in the presence of MG132. In (G) to (K), Ponceau stainings are included for loading reference. All GR24 treatments used a 5 μM concentration unless indicated.